Chimeric antigen and t cell receptors and methods of use

ABSTRACT

The invention provides a chimeric antigen receptor (CAR) or a T cell receptor (TCR) comprising extracellular domain disclosed herein. Some aspects of the invention relate to a polynucleotide encoding a chimeric antigen receptor (CAR) or a T cell receptor (TCR) comprising the extracellular domain disclosed herein. Other aspects of the invention relate to cells comprising the CAR or the TCR and their use in a T cell therapy.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/317,258, filed Apr. 1, 2016, which is herebyincorporated by reference in its entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Mar. 30, 2017, isnamed K-1031_02_SL.txt and is 414,963 bytes in size.

BACKGROUND OF THE INVENTION

Human cancers are by their nature comprised of normal cells that haveundergone a genetic or epigenetic conversion to become abnormal cancercells. In doing so, cancer cells begin to express proteins and otherantigens that are distinct from those expressed by normal cells. Theseaberrant tumor antigens can be used by the body's innate immune systemto specifically target and kill cancer cells. However, cancer cellsemploy various mechanisms to prevent immune cells, such as T and Blymphocytes, from successfully targeting cancer cells.

Current therapies T cell therapies rely on enriched or modified human Tcells to target and kill cancer cells in a patient. To increase theability of T cells to target and kill a particular cancer cell, methodshave been developed to engineer T cells to express constructs whichdirect T cells to a particular target cancer cell. Chimeric antigenreceptors (CARs) and engineered T cell receptors (TCRs), which comprisebinding domains capable of interacting with a particular tumor antigen,allow T cells to target and kill cancer cells that express theparticular tumor antigen.

A need exists for improved CARs and TCRs for targeting and killingcancer cells.

SUMMARY OF THE INVENTION

The present invention addresses this need by providing compositions andmethods comprising genetically engineered immune cells that expressantigen receptors (CARs) or T cell receptors (TCRs) which specificallytarget and kill cancer cells.

A CAR may comprise, for example, (i) an antigen-specific component(“antigen binding molecule”), (ii) one or more costimulatory domains(which includes a hinge domain), and (iii) one or more activatingdomains. Each domain may be heterogeneous, that is, comprised ofsequences derived from different protein chains. CAR-expressing immunecells (such as T cells) may be used in various therapies, includingcancer therapies.

As described in more detail below, including the Examples section, CARscomprising a costimulatory domain which includes a truncated hingedomain (“THD”) provides unexpectedly superior properties when comparedto a CAR comprising a costimulatory domain which includes a completehinge domain (“CHD”). Polynucleotides encoding such CARs can betransduced into T cells and the CARs are expressed in T cells, e.g., apatient's own T cells. When the transduced T cells are transplanted backto a patient, the CARS direct the T cells to recognize and bind anepitope present on the surface of cancer cells, thus, allowing bindingof cancer cells rather than non-cancerous cells. This binding leads toactivation of cytolytic mechanisms in the T cell that specifically killthe bound cancer cells. Prior to the present invention, it was unknownthat a CARs comprising a THD is superior to a CAR comprising a CHD.Thus, the present invention satisfies an unmet need that exists fornovel and improved therapies for treating cancer.

An aspect of the present invention is an isolated polynucleotideencoding a chimeric antigen receptor (CAR) or a T cell receptor (TCR),which comprises (i) an antigen binding molecule, (ii) a costimulatorydomain, and (iii) an activating domain. The costimulatory domain maycomprise an extracellular domain, a transmembrane domain, and anintracellular domain, wherein the extracellular domain comprises atruncated hinge domain consisting essentially of or consisting of (i) anamino acid sequence at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 96%, at least about 97%,at least about 98%, at least about 99%, or about 100% identical to aminoacids 123 to 152 of SEQ ID NO: 1 and, optionally, (ii) one to six aminoacids.

In some embodiments, the one to six amino acids are heterologous aminoacids.

In some embodiments, the truncated hinge domain consists essentially ofor consists of an amino acid sequence at least about 80%, at least about85%, at least about 90%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or about 100%identical to amino acids 123 to 152 of SEQ ID NO: 1.

In some embodiments, the amino acid sequence is encoded by a nucleotidesequence at least about 60%, at least about 70%, at least about 80%, atleast about 85%, at least about 90%, at least about 95%, at least about96%, at least about 97%, at least about 98%, at least about 99%, orabout 100% identical to SEQ ID NO: 2.

In some embodiments, the transmembrane domain is a transmembrane domainof 4-1BB/CD137, an alpha chain of a T cell receptor, a beta chain of a Tcell receptor, CD3 epsilon, CD4, CD5, CD8 alpha, CD9, CD16, CD19, CD22,CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137, CD154, or a zeta chainof a T cell receptor, or any combination thereof.

In some embodiments, the transmembrane domain comprises an amino acidsequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 5.

In some embodiments, the transmembrane domain is encoded by a nucleotidesequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 4.

In some embodiments, the intracellular domain comprises a signalingregion of 4-1BB/CD137, activating NK cell receptors, B7-H3, BAFFR, BLAME(SLAMF8), BTLA, CD100 (SEMA4D), CD103, CD160 (BY55), CD18, CD19, CD19a,CD2, CD247, CD27, CD276 (B7-H3), CD29, CD3 delta, CD3 epsilon, CD3gamma, CD30, CD4, CD40, CD49a, CD49D, CD49f, CD69, CD7, CD84, CD8alpha,CD8beta, CD96 (Tactile), CD11a, CD11b, CD11c, CD11d, CDS, CEACAM1, CRTAM, cytokine receptors, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS,GITR, HVEM (LIGHTR), IA4, ICAM-1, ICAM-1, Ig alpha (CD79a), IL2R beta,IL2R gamma, IL7R alpha, Immunoglobulin-like proteins, inducible T cellcostimulator (ICOS), integrins, ITGA4, ITGA4, ITGA6, ITGAD, ITGAE,ITGAL, ITGAM, ITGAX, ITGB2, ITGB7, ITGB1, KIRDS2, LAT, LFA-1, LFA-1, aligand that specifically binds with CD83, LIGHT, LIGHT (tumor necrosisfactor superfamily member 14; TNFSF14), LTBR, Ly9 (CD229), lymphocytefunction-associated antigen-1 (LFA-1 (CD11a/CD18), MHC class I molecule,NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), OX-40, PAG/Cbp,programmed death-1 (PD-1), PSGL1, SELPLG (CD162), signaling lymphocyticactivation molecules (SLAM proteins), SLAM (SLAMF1; CD150; IPO-3),SLAMF4 (CD244; 2B4), SLAMF6 (NTB-A; Ly108), SLAMF7, SLP-76, TNF receptorproteins, TNFR2, a Toll ligand receptor, TRANCE/RANKL, VLA1, or VLA-6,or a combination thereof.

In some embodiments, the intracellular domain comprises a 4-1BB/CD137signaling region.

In some embodiments, the intracellular domain comprises an amino acidsequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 7.

In some embodiments, the intracellular domain comprises an amino acidsequence encoded by a nucleotide sequence at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, at least about 99%, or about100% identical to SEQ ID NO: 6.

In some embodiments, the antigen binding molecule comprises a heavychain variable region (VH) and a light chain variable region (VL),wherein the VH comprises 3 complementarity determining regions (CDRs)and the VL comprises 3 CDRs.

In some embodiments, the antigen binding molecule specifically binds anantigen selected from the group consisting of 5T4, alphafetoprotein, Bcell maturation antigen (BCMA), CA-125, carcinoembryonic antigen, CD19,CD20, CD22, CD23, CD30, CD33, CD56, CD123, CD138, c-Met, CSPG4, C-typelectin-like molecule 1 (CLL-1), EGFRvIII, epithelial tumor antigen,ERBB2, FLT3, folate binding protein, GD2, GD3, HER1-HER2 in combination,HER2-HER3 in combination, HER2/Neu, HERV-K, HIV-1 envelope glycoproteingp41, HIV-1 envelope glycoprotein gp120, IL-11Ralpha, kappa chain,lambda chain, melanoma-associated antigen, mesothelin, MUC-1, mutatedp53, mutated ras, prostate-specific antigen, ROR1, or VEGFR2, or acombination thereof.

In some embodiments, the antigen binding molecule specifically bindsBCMA, CLL-1, or FLT3.

In some embodiments, the activation domain comprises an amino acidsequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 9 or SEQID NO: 251.

In some embodiments, the activation domain is encoded by a nucleotidesequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 8.

In some embodiments, the CAR or TCR further comprises a leader peptide.

In some embodiments, the leader peptide comprises an amino acid sequenceat least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, or about 100% identical to SEQ ID NO: 11.

In some embodiments, the leader peptide is encoded by a nucleotidesequence at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to SEQ ID NO: 10.

Another aspect of the present invention is a vector comprising thepolynucleotide of an above aspect or embodiment.

In some embodiments, the vector is an adenoviral vector, anadenovirus-associated vector, a DNA vector, a lentiviral vector, aplasmid, a retroviral vector, or an RNA vector, or any combinationthereof.

Yet another aspect of the present invention is a polypeptide encoded bythe polynucleotide of an above aspect or embodiment or the vector of anabove aspect or embodiment.

In another aspect, the present invention is a cell comprising thepolynucleotide of an above aspect or embodiment, the vector of an aboveaspect or embodiment, or the polypeptide of an above aspect orembodiment, or any combination thereof.

In some embodiments, the cell is a T cell.

In some embodiments, the T cell is an allogeneic T cell, an autologous Tcell, an engineered autologous T cell (eACT™), or a tumor-infiltratinglymphocyte (TIL).

In some embodiments, the T cell is a CD4+ T cell.

In some embodiments, the T cell is a CD8+ T cell.

In some embodiments, the T cell is an in vitro cell.

In some embodiments, the T cell is an autologous T cell.

An aspect of the present invention is a composition comprising thepolynucleotide of an above aspect or embodiment, comprising the vectorof an above aspect or embodiment, comprising the polypeptide of an aboveaspect or embodiment, or comprising the cell of an above aspect orembodiment.

In some embodiments, the composition is formulated to be delivered to asubject, optionally, comprising at least one pharmaceutically-acceptableexcipient.

Another aspect of the present invention is a method of making a cellexpressing a CAR or TCR comprising transducing a cell with thepolynucleotide of an above aspect or embodiment under suitableconditions.

In some embodiments, the method further comprises isolating the cell.

Yet another aspect of the present invention is a method of inducing animmunity against a tumor comprising administering to a subject aneffective amount of a cell comprising the polynucleotide of an aboveaspect or embodiment, comprising the vector of an above aspect orembodiment, or the polypeptide of an above aspect or embodiment, or anycombination thereof.

In another aspect, the present invention is a method of treating acancer in a subject in need thereof comprising administering to thesubject the polynucleotide of an above aspect or embodiment, the vectorof an above aspect or embodiment, the polypeptide of an above aspect orembodiment, the cell of an above aspect or embodiment, or thecomposition of an above aspect or embodiment.

In some embodiments, the cancer is a hematologic cancer.

In some embodiments, the cancer is of the white blood cells.

In some embodiments, the cancer is of the plasma cells.

In some embodiments, the cancer is leukemia, lymphoma, or myeloma.

In some embodiments, the cancer is acute lymphoblastic leukemia (ALL)(including non T cell ALL), acute myeloid leukemia, B cellprolymphocytic leukemia, B-cell acute lymphoid leukemia (“BALL”),blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma,chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML),chronic myeloid leukemia, chronic or acute leukemia, diffuse large Bcell lymphoma (DLBCL), follicular lymphoma (FL), hairy cell leukemia,Hodgkin's Disease, malignant lymphoproliferative conditions, MALTlymphoma, mantle cell lymphoma, Marginal zone lymphoma, monoclonalgammapathy of undetermined significance (MGUS), multiple myeloma,myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma(NHL), plasma cell proliferative disorder (including asymptomaticmyeloma (smoldering multiple myeloma or indolent myeloma), plasmablasticlymphoma, plasmacytoid dendritic cell neoplasm, plasmacytomas (includingplasma cell dyscrasia; solitary myeloma; solitary plasmacytoma;extramedullary plasmacytoma; and multiple plasmacytoma), POEMS syndrome(also known as Crow-Fukase syndrome; Takatsuki disease; and PEPsyndrome), primary mediastinal large B cell lymphoma (PMBC), small cell-or a large cell-follicular lymphoma, splenic marginal zone lymphoma(SMZL), systemic amyloid light chain amyloidosis, T-cell acute lymphoidleukemia (“TALL”), T-cell lymphoma, transformed follicular lymphoma, orWaldenstrom macroglobulinemia, or a combination thereof.

Generally, the present invention relates to Engineered Autologous CellTherapy, abbreviated as “eACT™,” also known as adoptive cell transfer.eACT™, is a process by which a patient's own T cells are collected andsubsequently genetically engineered to recognize and target one or moreantigens expressed on the cell surface of one or more specific cancercells. T cells may be engineered to express, for example, a CAR or TCR.CAR positive (CAR+) T cells are engineered to express a CAR. CARs maycomprise, e.g., an extracellular single chain variable fragment (scFv)with specificity for a particular tumor antigen, which is directly orindirectly linked to an intracellular signaling part comprising at leastone costimulatory domain, which is directly or indirectly linked to atleast one activating domain; the components may be arranged in anyorder. The costimulatory domain may be derived from a costimulatoryprotein known in the art, e.g., SEQ ID NO: 1, and the activating domainmay be derived from, e.g., any form of CD3-zeta. In some embodiments,the CAR is designed to have two, three, four, or more costimulatorydomains. In some embodiments, a CAR is engineered such that thecostimulatory domain is expressed as a separate polypeptide chain.Examples of CAR T cell therapies and constructs are described in U.S.Patent Publication Nos. 2013/0287748, 2014/0227237, 2014/0099309, and2014/0050708; International Patent Publications Nos. WO2012033885,WO2012079000, WO2014127261, WO2014186469, WO2015080981, WO2015142675,WO2016044745, and WO2016090369; and Sadelain et al, Cancer Discovery, 3:388-398 (2013), each of which is incorporated by reference in itsentirety.

Any aspect or embodiment described herein may be combined with any otheraspect or embodiment as disclosed herein. While the present inventionhas been described in conjunction with the detailed description thereof,the foregoing description is intended to illustrate and not limit thescope of the present invention, which is defined by the scope of theappended claims. Other aspects, advantages, and modifications are withinthe scope of the following claims.

The patent and scientific literature referred to herein establishes theknowledge that is available to those with skill in the art. All UnitedStates patents and published or unpublished United States patentapplications cited herein are incorporated by reference. All publishedforeign patents and patent applications cited herein are herebyincorporated by reference. All other published references, dictionaries,documents, manuscripts and scientific literature cited herein are herebyincorporated by reference.

Other features and advantages of the invention will be apparent from theDrawings and the following Detailed Description, including the Examples,and the claims.

BRIEF DESCRIPTION OF THE FIGURES

The above and further features will be more clearly appreciated from thefollowing detailed description when taken in conjunction with theaccompanying drawings. The drawings however are for illustrationpurposes only; not for limitation.

FIG. 1A shows a costimulatory protein having the amino acid sequence ofSEQ ID NO: 1. The costimulatory protein's hinge domain (solidunderline), transmembrane domain (dotted underline), and signalingdomain (dashed underline) are labeled. A novel truncated hinge domain(“THD”) is bolded. FIGS. 1B and 1C provide ribbon diagrams of theextracellular domain of the costimulatory protein having the amino acidsequence of SEQ ID NO: 1. FIG. 1B shows an example of a region withinthe amino acid sequence of SEQ ID NO: 1 used to derive one embodiment ofa hinge region in the context of CAR, i.e., a region containing aminoacids 114 to 152 of SEQ ID NO: 1 (herein referred to as a complete hingedomain or “CHD”; it is marked in black and dark grey). FIG. 1C shows theTHD which contain amino acids 123 to 152 of SEQ ID NO: 1 (marked inblack). In FIG. 1B, the portion of the hinge region that is excludedfrom FIG. 1C is marked dark grey and circled.

FIGS. 2A-2H show CLUTSTAL W (2.83) multiple sequence alignments of eightexample binding molecules disclosed herein. FIG. 2A shows a sequencealignment of example anti-CLL-1 binding molecules comprising a VHdomain. CDRs and framework regions FRs are shown, as determined byChothia numbering (FIG. 2A). FIG. 2B is a table providing the SEQ ID NOfor each VH and CDR illustrated in FIG. 2A. FIG. 2C shows a sequencealignment of example anti-CLL-1 binding molecules comprising a VLdomain. CDRs and FRs are shown, as determined by Chothia numbering (FIG.2C). FIG. 2D is a table providing the SEQ ID NO for each VH and CDRsequence illustrated in FIG. 2C. FIG. 2E shows a sequence alignment ofexample anti-BCMA binding molecules comprising a VH domain.Complementarity determining regions (CDRs) and framework regions (FRs)are shown, as determined by Chothia numbering (FIG. 2E). FIG. 2F is atable providing the SEQ ID NO for each VH and CDR illustrated in FIG.2E. FIG. 2G shows a sequence alignment of example anti-BCMA bindingmolecules comprising a VL domain. CDRs and FRs are shown, as determinedby Chothia numbering (FIG. 2G). FIG. 2H is a table providing the SEQ IDNO for each VH and CDR sequence illustrated in FIG. 2G.

FIG. 3 depicts CAR expression in primary human T cells electroporatedwith mRNA encoding for various CARs. Data obtained from CAR having acomplete hinge domain (“CHD”) is shown and data obtained from CAR havinga truncated hinge domain (“THD”) is shown.

FIGS. 4A-4X show IFNγ, IL-2, and TNFα production by electroporatedanti-FLT3 CART cells following 16 hours of co-culture with the indicatedtarget cell lines. FIGS. 4A-4B, 4G-4H, 4M-4N, and 4S-4T show IFNγproduction following co-culture with Namalwa, EoL-1, HL60, and MV4;11target cells, respectively. FIGS. 4C-4D, 4I-4J, 40-4P, and 4U-4V showIL-2 production following co-culture with Namalwa, EoL-1, HL60, andMV4;11 target cells, respectively. FIGS. 4E-4F, 4K-4L, 4Q-4R, and 4W-4Xshow TNFα production following co-culture with Namalwa, EoL-1, HL60, andMV4;11 target cells, respectively.

FIGS. 5A-5H show cytolytic activity of electroporated anti-FLT3 CAR Tcells against Namalwa (FIGS. 5A-5B), EoL1 (FIGS. 5C-5D), HL60 (FIGS.5E-5F), and MV4;11 (FIGS. 5G-5H) target cell lines following 16 hours ofco-culture.

FIGS. 6A-6B depict CAR expression in lentivirus transduced primary humanT cells from two healthy donors.

FIGS. 7A-7F show IFNγ (FIGS. 7A-7B), TNFα (FIGS. 7C-7D), and IL-2 (FIGS.7E-7F) production by lentivirus transduced CAR T cells from two healthydonors following 16 hours of co-culture with the indicated target celllines.

FIGS. 8A-8D show the average cytolytic activity over time from twohealthy donors expressing the anti-FLT3 CAR constructs co-cultured withNamalwa (FIG. 8A), EoL1 (FIG. 8B), MV4;11 (FIG. 8C), and HL60 (FIG. 8D)target cell lines for 16, 40, 64, 88, or 112 hours.

FIGS. 9A-9B depict proliferation of CFSE-labeled lentivirus transducedCAR T cells from two healthy donors following 5 days of co-culture withCD3-CD28 beads or the indicated target cell lines.

FIGS. 10A-10D depict CAR expression in lentivirus transduced primaryhuman T cells used for in vivo studies. FIGS. 10E-10F show graphicalrepresentations of measured bioluminescence imaging of labeled acutemyeloid leukemia (AML) cells following intravenous injection of eithercontrol (mock) or anti-FLT3 CAR T cells (10E3-CHD, 10E3-THD, or 8B5-THD)in a xenogeneic model, performed in duplicate. FIG. 10G provides thep-values for the respective data points in FIG. 10E. FIGS. 10H-10K showsurvival curves of mice injected with mock or 10E3-CHD (FIG. 10H), mockor 10E3-THD (FIG. 10I), mock or 8B5-THD (FIG. 10J), or 10E3-THD or8B5-THD (FIG. 10K) CAR T cells.

FIGS. 11A-11B shows CLL-1 CAR expression determined by protein L 6 hourspost mRNA electroporation.

FIGS. 12A-12C show the results from a cytokine release assay fromdifferent CLL-1 CAR-T cell constructs 24 hours after mRNAelectroporation. IL-2 (FIG. 12A), IFNγ(FIG. 12B), and TNFα (FIG. 12C)production levels are shown for controls (target alone, mock, GFP, andCD19 CAR T cells) and anti-CLL-1 CAR T cells (24C1_HL-THD, 24C1_HL_CHD,24C8_HL-CHD, and 24C8_HL_THD) co-cultured with Namalwa, MV4;11, U937,HL60, and EoL-1 cells, as indicated.

FIGS. 13A-13E show cytolytic activity of different CLL-1 CAR-T cellconstructs 24 hours after mRNA electroporation. T cells electroporatedwith control constructs (mock, GFP, and CD19 CAR) or anti-CLL-1 CARconstructs (24C8_HL-CHD and 24C8_HL_THD) were co-cultured with Namalwa(FIG. 13A), MV;411 (FIG. 13B), EoL-1 (FIG. 13C), HL-60 (FIG. 13D), andU937 target cells, and the percent of specific lysis of each target cellline was determined.

FIGS. 14A-14C show the results from a cytokine release assay fromdifferent transduced anti-CLL-1 CAR T cells 16 hours after co-culturewith different cell lines. IFNγ (FIG. 14A), IL-2 (FIG. 14B), and TNFα(FIG. 14C) production levels are shown for controls (target alone andmock) and transduced anti-CLL-1 CAR T cells (10E3 THD and 24C1_LH_THD)co-cultured with Namalwa, HL-60, or MV4;11 target cells, as indicated.

FIGS. 15A-15C show cytolytic activity from anti-CLL-1 CAR T cells(C1_24C1_LH_THD) 16 hours and 40 hours after co-culture with Namalwa(FIG. 15A), MV4;11 (FIG. 15B), or HL-60 (FIG. 15C) target cells.

FIGS. 16A-16F shows IFNγ, TNFα, and IL-2 production by lentivirustransduced CART cells from two healthy donors following 16 hours ofco-cultured with EoL-1 (Black), NCI-H929 (light grey), or MM1S (grey)target cell lines. FIGS. 16A and 16B show the IFNγ (pg/ml; y-axis)production in lentivirus transduced CAR T cells from a first donor (FIG.6A) and a second donor (FIG. 16B). FIGS. 16C and 16D show the TNFα(pg/ml; y-axis) production in lentivirus transduced CAR T cells from afirst donor (FIG. 16C) and a second donor (FIG. 16D). FIGS. 16E and 16Fshow the IL-2 production (pg/ml; y-axis) in lentivirus transduced CAR Tcells from a first donor (FIG. 16E) and a second donor (FIG. 16F).

FIGS. 17A-17F show the average cytolytic activity (as a percentage ofviable target cells remaining; y-axis) over time from two healthy donorsexpressing the indicated CARs co-cultured with EoL1 (FIGS. 17A and 17B),NCI-H929 (FIGS. 17C and 17D), or MM1S (FIGS. 17E and 17F) target cellsfor 16 hours, 40 hours, 64 hours, 88 hours, or 112 hours. FIGS. 17A and17B show the average cytolytic activity of transduced CAR T cells from afirst donor (FIG. 17A) and a second donor (FIG. 17B) co-cultured withEoL1 target cells for 16 hours, 40 hours, 64 hours, 88 hours, or 112hours. FIGS. 17C and 17D show the average cytolytic activity oftransduced CAR T cells from a first donor (FIG. 17C) and a second donor(FIG. 17D) co-cultured with NCI-H929 target cells for 16 hours, 40hours, 64 hours, 88 hours, or 112 hours. FIGS. 17E and 17F show theaverage cytolytic activity of transduced CAR T cells from a first donor(FIG. 17E) and a second donor (FIG. 17F) co-cultured with MM1S targetcells for 16 hours, 40 hours, 64 hours, 88 hours, or 112 hours.

FIGS. 18A and 18B show proliferation of CFSE-labeled lentivirustransduced CAR T cells from a first healthy donor (FIG. 18A) and asecond healthy donor (FIG. 18B) following 6 days of co-culture withCD3-CD28 beads (top row), EoL-1 (second row), NCI-H929 (third row), orMM1S (bottom row) target cell lines.

FIG. 19A and FIG. 19B are graphs showing thermostability of chimericantigen receptors (CARs) of the present invention. FIG. 19A: In aphosphate-buffered saline (PBS) solution, a CAR comprising anextracellular domain with a truncated hinge domain (“THD”) has a highermelting temperature relative to a CAR comprising an extracellular domainwith a complete hinge domain (“CHD”). FIG. 19B: In the presence of 50 mMNaCl, a CAR comprising an extracellular domain with a THD has a highermelting temperature relative to a CAR comprising an extracellular domainwith a CHD.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to novel polypeptides comprising a noveltruncated hinge domain (“THD”) and polynucleotides encoding the same.Some aspects of the invention relate to a polynucleotide encoding achimeric antigen receptor (CAR) or a T cell receptor (TCR) comprisingthe THD disclosed herein. The present invention also provides vectors(e.g., viral vectors) comprising such polynucleotides and compositionscomprising such vectors. The present invention further providespolynucleotides encoding such CARs or TCRs and compositions comprisingsuch polynucleotides. The present invention additionally providesengineered cells (e.g., T cells) comprising such polynucleotides and/ortransduced with such viral vectors and compositions comprising suchengineered cells. The present invention provides compositions (e.g.,pharmaceutical compositions) including a plurality of engineered Tcells. The present invention provides methods for manufacturing suchengineered T cells and compositions and uses (e.g., in treating amelanoma) of such engineered T cells and compositions. And, the presentinvention provides a method of inducing an immunity against a tumorcomprising administering to a subject an effective amount of a cellcomprising a polynucleotide, a vector, or a polypeptide of the presentinvention. Other aspects of the invention relate to cells comprising theCAR or the TCR and their use in a T cell therapy, e.g., an autologouscell therapy (eACT™), for the treatment of a patient suffering from acancer.

Definitions

In order for the present invention to be more readily understood,certain terms are first defined below. Additional definitions for thefollowing terms and other terms are set forth throughout theSpecification.

As used in this Specification and the appended claims, the singularforms “a,” “an” and “the” include plural referents unless the contextclearly dictates otherwise.

Unless specifically stated or obvious from context, as used herein, theterm “or” is understood to be inclusive and covers both “or” and “and”.

The term “and/or” where used herein is to be taken as specificdisclosure of each of the two specified features or components with orwithout the other. Thus, the term “and/or” as used in a phrase such as“A and/or B” herein is intended to include A and B; A or B; A (alone);and B (alone). Likewise, the term “and/or” as used in a phrase such as“A, B, and/or C” is intended to encompass each of the following aspects:A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B andC; A (alone); B (alone); and C (alone).

The terms “e.g.,” and “i.e.” as used herein, are used merely by way ofexample, without limitation intended, and should not be construed asreferring only those items explicitly enumerated in the specification.

The terms “or more”, “at least”, “more than”, and the like, e.g., “atleast one” are understood to include but not be limited to at least 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150,200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 ormore than the stated value. Also included is any greater number orfraction in between.

Conversely, the term “no more than” includes each value less than thestated value. For example, “no more than 100 nucleotides” includes 100,99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82,81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64,63, 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46,45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28,27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10,9, 8, 7, 6, 5, 4, 3, 2, 1, and 0 nucleotides. Also included is anylesser number or fraction in between.

The terms “plurality”, “at least two”, “two or more”, “at least second”,and the like, are understood to include but not limited to at least 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109,110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137,138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200,300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more.Also included is any greater number or fraction in between.

Throughout the specification the word “comprising,” or variations suchas “comprises” or “comprising,” will be understood to imply theinclusion of a stated element, integer or step, or group of elements,integers or steps, but not the exclusion of any other element, integeror step, or group of elements, integers or steps. It is understood thatwherever aspects are described herein with the language “comprising,”otherwise analogous aspects described in terms of “consisting of” and/or“consisting essentially of” are also provided.

Unless specifically stated or evident from context, as used herein, theterm “about” refers to a value or composition that is within anacceptable error range for the particular value or composition asdetermined by one of ordinary skill in the art, which will depend inpart on how the value or composition is measured or determined, i.e.,the limitations of the measurement system. For example, “about” or“comprising essentially of” can mean within one or more than onestandard deviation per the practice in the art. “About” or “comprisingessentially of” can mean a range of up to 10% (i.e., ±10%). Thus,“about” can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%,2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% greater or less than thestated value. For example, about 5 mg can include any amount between 4.5mg and 5.5 mg. Furthermore, particularly with respect to biologicalsystems or processes, the terms can mean up to an order of magnitude orup to 5-fold of a value. When particular values or compositions areprovided in the instant disclosure, unless otherwise stated, the meaningof “about” or “comprising essentially of” should be assumed to be withinan acceptable error range for that particular value or composition.

As described herein, any concentration range, percentage range, ratiorange or integer range is to be understood to be inclusive of the valueof any integer within the recited range and, when appropriate, fractionsthereof (such as one-tenth and one-hundredth of an integer), unlessotherwise indicated.

Units, prefixes, and symbols used herein are provided using theirSystème International de Unites (SI) accepted form. Numeric ranges areinclusive of the numbers defining the range.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure is related. For example, Juo, “TheConcise Dictionary of Biomedicine and Molecular Biology”, 2^(nd) ed.,(2001), CRC Press; “The Dictionary of Cell & Molecular Biology”, 5^(th)ed., (2013), Academic Press; and “The Oxford Dictionary Of BiochemistryAnd Molecular Biology”, Cammack et al. eds., 2^(nd) ed, (2006), OxfordUniversity Press, provide those of skill in the art with a generaldictionary for many of the terms used in this disclosure.

“Administering” refers to the physical introduction of an agent to asubject, using any of the various methods and delivery systems known tothose skilled in the art. Exemplary routes of administration for theformulations disclosed herein include intravenous, intramuscular,subcutaneous, intraperitoneal, spinal or other parenteral routes ofadministration, for example by injection or infusion. The phrase“parenteral administration” as used herein means modes of administrationother than enteral and topical administration, usually by injection, andincludes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intralymphatic, intralesional, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal,epidural and intrasternal injection and infusion, as well as in vivoelectroporation. In some embodiments, the formulation is administeredvia a non-parenteral route, e.g., orally. Other non-parenteral routesinclude a topical, epidermal or mucosal route of administration, forexample, intranasally, vaginally, rectally, sublingually or topically.Administering can also be performed, for example, once, a plurality oftimes, and/or over one or more extended periods.

The term “antibody” (Ab) includes, without limitation, a glycoproteinimmunoglobulin which binds specifically to an antigen. In general, andantibody can comprise at least two heavy (H) chains and two light (L)chains interconnected by disulfide bonds, or an antigen-binding moleculethereof. Each H chain comprises a heavy chain variable region(abbreviated herein as VH) and a heavy chain constant region. The heavychain constant region comprises three constant domains, CH1, CH2 andCH3. Each light chain comprises a light chain variable region(abbreviated herein as VL) and a light chain constant region. The lightchain constant region is comprises one constant domain, CL. The VH andVL regions can be further subdivided into regions of hypervariability,termed complementarity determining regions (CDRs), interspersed withregions that are more conserved, termed framework regions (FR). Each VHand VL comprises three CDRs and four FRs, arranged from amino-terminusto carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, and FR4. The variable regions of the heavy and light chainscontain a binding domain that interacts with an antigen. The constantregions of the Abs may mediate the binding of the immunoglobulin to hosttissues or factors, including various cells of the immune system (e.g.,effector cells) and the first component (C1q) of the classicalcomplement system.

Antibodies can include, for example, monoclonal antibodies,recombinantly produced antibodies, monospecific antibodies,multispecific antibodies (including bispecific antibodies), humanantibodies, engineered antibodies, humanized antibodies, chimericantibodies, immunoglobulins, synthetic antibodies, tetrameric antibodiescomprising two heavy chain and two light chain molecules, an antibodylight chain monomer, an antibody heavy chain monomer, an antibody lightchain dimer, an antibody heavy chain dimer, an antibody lightchain-antibody heavy chain pair, intrabodies, antibody fusions(sometimes referred to herein as “antibody conjugates”), heteroconjugateantibodies, single domain antibodies, monovalent antibodies, singlechain antibodies or single-chain Fvs (scFv), camelized antibodies,affybodies, Fab fragments, F(ab′)₂ fragments, disulfide-linked Fvs(sdFv), anti-idiotypic (anti-Id) antibodies (including, e.g.,anti-anti-Id antibodies), minibodies, domain antibodies, syntheticantibodies (sometimes referred to herein as “antibody mimetics”), andantigen-binding fragments of any of the above. In certain embodiments,antibodies described herein refer to polyclonal antibody populations.

An immunoglobulin may derive from any of the commonly known isotypes,including but not limited to IgA, secretory IgA, IgG, IgE and IgM. IgGsubclasses are also well known to those in the art and include but arenot limited to human IgG1, IgG2, IgG3 and IgG4. “Isotype” refers to theAb class or subclass (e.g., IgM or IgG1) that is encoded by the heavychain constant region genes. The term “antibody” includes, by way ofexample, both naturally occurring and non-naturally occurring Abs;monoclonal and polyclonal Abs; chimeric and humanized Abs; human ornonhuman Abs; wholly synthetic Abs; and single chain Abs. A nonhuman Abmay be humanized by recombinant methods to reduce its immunogenicity inman. Where not expressly stated, and unless the context indicatesotherwise, the term “antibody” also includes an antigen-binding fragmentor an antigen-binding portion of any of the aforementionedimmunoglobulins, and includes a monovalent and a divalent fragment orportion, and a single chain Ab.

An “antigen binding molecule,” “antigen binding portion,” or “antibodyfragment” refers to any molecule that comprises the antigen bindingparts (e.g., CDRs) of the antibody from which the molecule is derived.An antigen binding molecule can include the antigenic complementaritydetermining regions (CDRs). Examples of antibody fragments include, butare not limited to, Fab, Fab′, F(ab′)2, and Fv fragments, dAb, linearantibodies, scFv antibodies, and multispecific antibodies formed fromantigen binding molecules. Peptibodies (i.e., Fc fusion moleculescomprising peptide binding domains) are another example of suitableantigen binding molecules. In some embodiments, the antigen bindingmolecule binds to an antigen on a tumor cell. In some embodiments, theantigen binding molecule binds to an antigen on a cell involved in ahyperproliferative disease or to a viral or bacterial antigen. Incertain embodiments, the antigen binding molecule binds to BCMA, CLL-1,or FLT3. In further embodiments, the antigen binding molecule is anantibody fragment that specifically binds to the antigen, including oneor more of the complementarity determining regions (CDRs) thereof. Infurther embodiments, the antigen binding molecule is a single chainvariable fragment (scFv). In some embodiments, the antigen bindingmolecule comprises or consists of avimers.

As used herein, the term “variable region” or “variable domain” is usedinterchangeably and are common in the art. The variable region typicallyrefers to a portion of an antibody, generally, a portion of a light orheavy chain, typically about the amino-terminal 110 to 120 amino acidsin the mature heavy chain and about 90 to 115 amino acids in the maturelight chain, which differ extensively in sequence among antibodies andare used in the binding and specificity of a particular antibody for itsparticular antigen. The variability in sequence is concentrated in thoseregions called complementarity determining regions (CDRs) while the morehighly conserved regions in the variable domain are called frameworkregions (FR). Without wishing to be bound by any particular mechanism ortheory, it is believed that the CDRs of the light and heavy chains areprimarily responsible for the interaction and specificity of theantibody with antigen. In certain embodiments, the variable region is ahuman variable region. In certain embodiments, the variable regioncomprises rodent or murine CDRs and human framework regions (FRs). Inparticular embodiments, the variable region is a primate (e.g.,non-human primate) variable region. In certain embodiments, the variableregion comprises rodent or murine CDRs and primate (e.g., non-humanprimate) framework regions (FRs).

The terms “VL” and “VL domain” are used interchangeably to refer to thelight chain variable region of an antibody or an antigen-bindingmolecule thereof.

The terms “VH” and “VH domain” are used interchangeably to refer to theheavy chain variable region of an antibody or an antigen-bindingmolecule thereof.

A number of definitions of the CDRs are commonly in use: Kabatnumbering, Chothia numbering, AbM numbering, or contact numbering. TheAbM definition is a compromise between the two used by OxfordMolecular's AbM antibody modelling software. The contact definition isbased on an analysis of the available complex crystal structures.

TABLE 1 CDR Numbering Loop Kabat AbM Chothia Contact L1 L24--L34L24--L34 L24--L34 L30--L36 L2 L50--L56 L50--L56 L50--L56 L46--L55 L3L89--L97 L89--L97 L89--L97 L89--L96 H1 H31--H35B H26--H35B H26--H32..34H30--H35B (Kabat Numbering) H1 H31--H35 H26--H35 H26--H32 H30--H35(Chothia Numbering) H2 H50--H65 H50--H58 H52--H56 H47--H58 H3 H95--H102H95--H102 H95--H102 H93--H101

The term “Kabat numbering” and like terms are recognized in the art andrefer to a system of numbering amino acid residues in the heavy andlight chain variable regions of an antibody, or an antigen-bindingmolecule thereof. In certain aspects, the CDRs of an antibody can bedetermined according to the Kabat numbering system (see, e.g., Kabat E A& Wu T T (1971) Ann NY Acad Sci 190: 382-391 and Kabat E A et al.,(1991) Sequences of Proteins of Immunological Interest, Fifth Edition,U.S. Department of Health and Human Services, NIH Publication No.91-3242). Using the Kabat numbering system, CDRs within an antibodyheavy chain molecule are typically present at amino acid positions 31 to35, which optionally can include one or two additional amino acids,following 35 (referred to in the Kabat numbering scheme as 35A and 35B)(CDR1), amino acid positions 50 to 65 (CDR2), and amino acid positions95 to 102 (CDR3). Using the Kabat numbering system, CDRs within anantibody light chain molecule are typically present at amino acidpositions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), andamino acid positions 89 to 97 (CDR3). In a specific embodiment, the CDRsof the antibodies described herein have been determined according to theKabat numbering scheme.

In certain aspects, the CDRs of an antibody can be determined accordingto the Chothia numbering scheme, which refers to the location ofimmunoglobulin structural loops (see, e.g., Chothia C & Lesk A M,(1987), J Mol Biol 196: 901-917; Al-Lazikani B et al., (1997) J Mol Biol273: 927-948; Chothia C et al., (1992) J Mol Biol 227: 799-817;Tramontano A et al., (1990) J Mol Biol 215(1): 175-82; and U.S. Pat. No.7,709,226). Typically, when using the Kabat numbering convention, theChothia CDR-H1 loop is present at heavy chain amino acids 26 to 32, 33,or 34, the Chothia CDR-H2 loop is present at heavy chain amino acids 52to 56, and the Chothia CDR-H3 loop is present at heavy chain amino acids95 to 102, while the Chothia CDR-L1 loop is present at light chain aminoacids 24 to 34, the Chothia CDR-L2 loop is present at light chain aminoacids 50 to 56, and the Chothia CDR-L3 loop is present at light chainamino acids 89 to 97. The end of the Chothia CDR-HI loop when numberedusing the Kabat numbering convention varies between H32 and H34depending on the length of the loop (this is because the Kabat numberingscheme places the insertions at H35A and H35B; if neither 35A nor 35B ispresent, the loop ends at 32; if only 35A is present, the loop ends at33; if both 35A and 35B are present, the loop ends at 34). In a specificembodiment, the CDRs of the antibodies described herein have beendetermined according to the Chothia numbering scheme.

As used herein, the terms “constant region” and “constant domain” areinterchangeable and have a meaning common in the art. The constantregion is an antibody portion, e.g., a carboxyl terminal portion of alight and/or heavy chain which is not directly involved in binding of anantibody to antigen but which can exhibit various effector functions,such as interaction with the Fc receptor. The constant region of animmunoglobulin molecule generally has a more conserved amino acidsequence relative to an immunoglobulin variable domain.

As used herein, the term “heavy chain” when used in reference to anantibody can refer to any distinct type, e.g., alpha (α), delta (δ),epsilon (ε), gamma (γ) and mu (μ), based on the amino acid sequence ofthe constant domain, which give rise to IgA, IgD, IgE, IgG and IgMclasses of antibodies, respectively, including subclasses of IgG, e.g.,IgG₁, IgG₂, IgG₃ and IgG₄.

As used herein, the term “light chain” when used in reference to anantibody can refer to any distinct type, e.g., kappa (κ) or lambda (λ)based on the amino acid sequence of the constant domains. Light chainamino acid sequences are well known in the art. In specific embodiments,the light chain is a human light chain.

“Binding affinity” generally refers to the strength of the sum total ofnon-covalent interactions between a single binding site of a molecule(e.g., an antibody) and its binding partner (e.g., an antigen). Unlessindicated otherwise, as used herein, “binding affinity” refers tointrinsic binding affinity which reflects a 1:1 interaction betweenmembers of a binding pair (e.g., antibody and antigen). The affinity ofa molecule X for its partner Y can generally be represented by thedissociation constant (K_(D)). Affinity can be measured and/or expressedin a number of ways known in the art, including, but not limited to,equilibrium dissociation constant (K_(D)), and equilibrium associationconstant (K_(A)). The K_(D) is calculated from the quotient ofk_(off)/k_(on), whereas K_(A) is calculated from the quotient ofk_(on)/k_(off). k_(on) refers to the association rate constant of, e.g.,an antibody to an antigen, and k_(off) refers to the dissociation of,e.g., an antibody to an antigen. The k_(on) and k_(off) can bedetermined by techniques known to one of ordinary skill in the art, suchas BIACORE® or KinExA.

As used herein, a “conservative amino acid substitution” is one in whichthe amino acid residue is replaced with an amino acid residue having asimilar side chain. Families of amino acid residues having side chainshave been defined in the art. These families include amino acids withbasic side chains (e.g., lysine, arginine, histidine), acidic sidechains (e.g., aspartic acid, glutamic acid), uncharged polar side chains(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine,leucine, isoleucine, proline, phenylalanine, methionine), beta-branchedside chains (e.g., threonine, valine, isoleucine) and aromatic sidechains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Incertain embodiments, one or more amino acid residues within a CDR(s) orwithin a framework region(s) of an antibody or antigen-binding moleculethereof can be replaced with an amino acid residue with a similar sidechain.

As, used herein, the term “heterologous” means from any source otherthan naturally occurring sequences. For example, a heterologous sequenceincluded as a part of a costimulatory protein having the amino acidsequence of SEQ ID NO: 1, e.g., the corresponding human costimulatoryprotein, is amino acids that do not naturally occur as, i.e., do notalign with, the wild type human costimulatory protein. For example, aheterologous nucleotide sequence refers to a nucleotide sequence otherthan that of the wild type human costimulatory protein-encodingsequence.

As used herein, an “epitope” is a term in the art and refers to alocalized region of an antigen to which an antibody can specificallybind. An epitope can be, for example, contiguous amino acids of apolypeptide (linear or contiguous epitope) or an epitope can, forexample, come together from two or more non-contiguous regions of apolypeptide or polypeptides (conformational, non-linear, discontinuous,or non-contiguous epitope). In certain embodiments, the epitope to whichan antibody binds can be determined by, e.g., NMR spectroscopy, X-raydiffraction crystallography studies, ELISA assays, hydrogen/deuteriumexchange coupled with mass spectrometry (e.g., liquid chromatographyelectrospray mass spectrometry), array-based oligo-peptide scanningassays, and/or mutagenesis mapping (e.g., site-directed mutagenesismapping). For X-ray crystallography, crystallization may be accomplishedusing any of the known methods in the art (e.g., Giegé R et al., (1994)Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A(1990) Eur J Biochem 189: 1-23; Chayen N E (1997) Structure 5:1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303).Antibody:antigen crystals may be studied using well known X-raydiffraction techniques and may be refined using computer software suchas X-PLOR (Yale University, 1992, distributed by Molecular Simulations,Inc.; see e.g. Meth Enzymol (1985) volumes 114 & 115, eds Wyckoff H W etal.; U.S. 2004/0014194), and BUSTER (Bricogne G (1993) Acta CrystallogrD Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A:361-423, ed Carter C W; Roversi P et al., (2000) Acta Crystallogr D BiolCrystallogr 56(Pt 10): 1316-1323). Mutagenesis mapping studies may beaccomplished using any method known to one of skill in the art. See,e.g., Champe M et al., (1995) J Biol Chem 270: 1388-1394 and CunninghamB C & Wells J A (1989) Science 244: 1081-1085 for a description ofmutagenesis techniques, including alanine scanning mutagenesistechniques.

As used herein, an antigen binding molecule, an antibody, or an antigenbinding molecule thereof “cross-competes” with a reference antibody oran antigen binding molecule thereof if the interaction between anantigen and the first binding molecule, an antibody, or an antigenbinding molecule thereof blocks, limits, inhibits, or otherwise reducesthe ability of the reference binding molecule, reference antibody, or anantigen binding molecule thereof to interact with the antigen. Crosscompetition can be complete, e.g., binding of the binding molecule tothe antigen completely blocks the ability of the reference bindingmolecule to bind the antigen, or it can be partial, e.g., binding of thebinding molecule to the antigen reduces the ability of the referencebinding molecule to bind the antigen. In certain embodiments, an antigenbinding molecule that cross-competes with a reference antigen bindingmolecule binds the same or an overlapping epitope as the referenceantigen binding molecule. In other embodiments, the antigen bindingmolecule that cross-competes with a reference antigen binding moleculebinds a different epitope as the reference antigen binding molecule.Numerous types of competitive binding assays can be used to determine ifone antigen binding molecule competes with another, for example: solidphase direct or indirect radioimmunoassay (MA); solid phase direct orindirect enzyme immunoassay (EIA); sandwich competition assay (Stahli etal., 1983, Methods in Enzymology 9:242-253); solid phase directbiotin-avidin EIA (Kirkland et al., 1986, J. Immunol. 137:3614-3619);solid phase direct labeled assay, solid phase direct labeled sandwichassay (Harlow and Lane, 1988, Antibodies, A Laboratory Manual, ColdSpring Harbor Press); solid phase direct label RIA using 1-125 label(Morel et al., 1988, Molec. Immunol. 25:7-15); solid phase directbiotin-avidin EIA (Cheung, et al., 1990, Virology 176:546-552); anddirect labeled RIA (Moldenhauer et al., 1990, Scand. J. Immunol.32:77-82).

As used herein, the terms “immunospecifically binds,”“immunospecifically recognizes,” “specifically binds,” and “specificallyrecognizes” are analogous terms in the context of antibodies and referto molecules that bind to an antigen (e.g., epitope or immune complex)as such binding is understood by one skilled in the art. For example, amolecule that specifically binds to an antigen may bind to otherpeptides or polypeptides, generally with lower affinity as determinedby, e.g., immunoassays, BIACORE®, KinExA 3000 instrument (SapidyneInstruments, Boise, Id.), or other assays known in the art. In aspecific embodiment, molecules that specifically bind to an antigen bindto the antigen with a K_(A) that is at least 2 logs, 2.5 logs, 3 logs, 4logs or greater than the K_(A) when the molecules bind to anotherantigen.

In another embodiment, molecules that specifically bind to an antigenbind with a dissociation constant (K_(d)) of about 1×10⁻⁷ M. In someembodiments, the antigen binding molecule specifically binds an antigenwith “high affinity” when the K_(d) is about 1×10⁻⁹ M to about 5×10⁻⁹ M.In some embodiments, the antigen binding molecule specifically binds anantigen with “very high affinity” when the K_(d) is 1×10⁻¹⁰ M to about5×10⁻¹⁰ M. In one embodiment, the antigen binding molecule has a K_(d)of 10⁻⁹ M. In one embodiment, the off-rate is less than about 1×10⁻⁵. Inother embodiments, the antigen binding molecule binds human BCMA with aK_(d) of between about 1×10⁻⁷ M and about 1×10⁻¹³ M. In yet anotherembodiment, the antigen binding molecule binds human BCMA with a K_(d)of about 1×10⁻¹⁰ M to about 5×10⁻¹⁰ M.

In a specific embodiment, provided herein is an antibody or an antigenbinding molecule thereof that binds to a target human antigen, e.g.,human BCMA or human CLL-1, with higher affinity than to another speciesof the target antigen, e.g., a non-human BCMA or a non-human CLL-1. Incertain embodiments, provided herein is an antibody or an antigenbinding molecule t thereof that binds to the target human antigen, e.g.,human BCMA or human CLL-1, with a 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70% or higher affinity than to another speciesof the target antigen as measured by, e.g., a radioimmunoassay, surfaceplasmon resonance, or kinetic exclusion assay. In a specific embodiment,an antibody or an antigen binding molecule thereof described herein,which binds to a target human antigen, will bind to another species ofthe target antigen with less than 10%, 15%, or 20% of the binding of theantibody or an antigen binding molecule thereof to the human antigen asmeasured by, e.g., a radioimmunoassay, surface plasmon resonance, orkinetic exclusion assay.

An “antigen” refers to any molecule that provokes an immune response oris capable of being bound by an antibody or an antigen binding molecule.The immune response may involve either antibody production, or theactivation of specific immunologically-competent cells, or both. Aperson of skill in the art would readily understand that anymacromolecule, including virtually all proteins or peptides, can serveas an antigen. An antigen can be endogenously expressed, i.e. expressedby genomic DNA, or can be recombinantly expressed. An antigen can bespecific to a certain tissue, such as a cancer cell, or it can bebroadly expressed. In addition, fragments of larger molecules can act asantigens. In one embodiment, antigens are tumor antigens. In oneparticular embodiment, the antigen is all or a fragment of BCMA, FLT3,or CLL-1.

The term “neutralizing” refers to an antigen binding molecule, scFv,antibody, or a fragment thereof, that binds to a ligand and prevents orreduces the biological effect of that ligand. In some embodiments, theantigen binding molecule, scFv, antibody, or a fragment thereof,directly blocking a binding site on the ligand or otherwise alters theligand's ability to bind through indirect means (such as structural orenergetic alterations in the ligand). In some embodiments, the antigenbinding molecule, scFv, antibody, or a fragment thereof prevents theprotein to which it is bound from performing a biological function.

As used herein, the term “BCMA” refers to B cell maturation antigen,which can include, but is not limited to, native BCMA, an isoform ofBCMA, or an interspecies BCMA homolog of BCMA. BCMA (also known asTNFRSF17, CD269, and TNFRSF13A) is a member of the tumor necrosis factor(TNF)-receptor superfamily. BCMA is expressed on the surface of multiplemyeloma cells, while highly restricted to plasma cells and a subset ofmature B cells in healthy tissue. The amino acid sequence of human BCMA(hBCMA) is provided in NCBI Accession Q02223.2 (GI:313104029). As usedherein, BCMA includes human BCMA and non-human BCMA homologs, as well asvariants, fragments, or post-transnationally modified forms thereof,including, but not limited to, N- and O-linked glycosylated forms ofBCMA. BCMA proteins may further include fragments comprising all or aportion of the extracellular domain of BCMA (e.g., all or a portion ofamino acids 1-54 of hBCMA).

As used herein, the term “CLL-1” refers to C-type lectin-likemolecule-1, which can include, but is not limited to native CLL-1, anisoform of CLL-1, or an interspecies CLL-1 homolog of CLL-1. CLL-1 (alsoknown as C-type lectin domain family 12 member A, CLEC12A, dendriticcell-associated lectin 2, DCAL-2, myeloid inhibitory C-type lectin-likereceptor, and MICL) is a cell surface receptor that modulates signalingcascades and mediates tyrosine phosphorylation of target MAP kinases.CLL-1 expression is observed, e.g., in acute myeloid leukemia (AML)cells. The amino acid sequence of human CLL-1 (hCLL-1) is provided inUniProtKB/Swiss-Prot Accession No. Q5QGZ9.3 (GI:308153619). As usedherein, CLL-1 includes human CLL-1 and non-human CLL-1 homologs, as wellas variants, fragments, or post-transnationally modified forms thereof,including, but not limited to, N- and O-linked glycosylated forms ofCLL-1.

As used herein the term “FLT3” refers to Fms-like tyrosine kinase 3(FLT-3), which can include, but is not limited to native FLT3, anisoform of FLT3, or an interspecies FLT3 homolog of FLT3. FLT3 (alsoknown as Cluster of differentiation antigen 135 (CD135), receptor-typetyrosine-protein kinase FLT3, FMS-related tyrosine kinase 3, stem celltyrosine kinase 1, FL cytokine receptor, growth factor receptor tyrosinekinase type III, STK1, or fetal liver kinase-2 (Flk2)) is a cytokinereceptor which belongs to the receptor tyrosine kinase class III. CD135is the receptor for the cytokine Flt3 ligand (FLT3L). FLT3 is expressedon the surface of various hematopoietic progenitor cells and on thesurface of acute myeloid leukemia (AML) cells. The amino acid sequenceof human FLT3 (hFLT3) is provided in UniProtKB/Swiss-Prot Accession No.P36888 (GI:156630887). As used herein, FLT3 includes human FLT3 andnon-human FLT3 homologs, as well as variants, fragments, orpost-transnationally modified forms thereof, including, but not limitedto, N- and O-linked glycosylated forms of FLT3.

The term “autologous” refers to any material derived from the sameindividual to which it is later to be re-introduced. For example, theengineered autologous cell therapy (eACT™) method described hereininvolves collection of lymphocytes from a patient, which are thenengineered to express, e.g., a CAR construct, and then administered backto the same patient.

The term “allogeneic” refers to any material derived from one individualwhich is then introduced to another individual of the same species,e.g., allogeneic T cell transplantation.

The terms “transduction” and “transduced” refer to the process wherebyforeign DNA is introduced into a cell via viral vector (see Jones etal., “Genetics: principles and analysis,” Boston: Jones & Bartlett Publ.(1998)). In some embodiments, the vector is a retroviral vector, a DNAvector, a RNA vector, an adenoviral vector, a baculoviral vector, anEpstein Barr viral vector, a papovaviral vector, a vaccinia viralvector, a herpes simplex viral vector, an adenovirus associated vector,a lentiviral vector, or any combination thereof.

As used herein, the term “truncated” refers to anything less than thewhole. For example, a truncated hinge domain (alternatively referred toherein as “THD”) amino acid sequence can include any amino acid sequenceshorter than the full length or complete hinge domain (“CHD”). In someembodiments, a THD consists essentially of or consists of amino acids118-152, 119-152, 120-152, 121-152, 122-152, 123-152, 124-152, 125-152,126-152, 127-152, 128-152, 129-152, or 130-152, of SEQ ID NO: 1. In oneembodiment, the THD consists essentially of or consists of the aminoacid sequence of SEQ ID NO: 3, which consists of amino acids 123 to 152of SEQ ID NO: 1.

A “cancer” refers to a broad group of various diseases characterized bythe uncontrolled growth of abnormal cells in the body. Unregulated celldivision and growth results in the formation of malignant tumors thatinvade neighboring tissues and may also metastasize to distant parts ofthe body through the lymphatic system or bloodstream. A “cancer” or“cancer tissue” can include a tumor. Examples of cancers that can betreated by the methods of the present invention include, but are notlimited to, cancers of the immune system including lymphoma, leukemia,myeloma, and other leukocyte malignancies. In some embodiments, themethods of the present invention can be used to reduce the tumor size ofa tumor derived from, for example, bone cancer, pancreatic cancer, skincancer, cancer of the head or neck, cutaneous or intraocular malignantmelanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of theanal region, stomach cancer, testicular cancer, uterine cancer,carcinoma of the fallopian tubes, carcinoma of the endometrium,carcinoma of the cervix, carcinoma of the vagina, carcinoma of thevulva, multiple myeloma, Hodgkin's Disease, non-Hodgkin's lymphoma(NHL), primary mediastinal large B cell lymphoma (PMBC), diffuse large Bcell lymphoma (DLBCL), follicular lymphoma (FL), transformed follicularlymphoma, splenic marginal zone lymphoma (SMZL), cancer of theesophagus, cancer of the small intestine, cancer of the endocrinesystem, cancer of the thyroid gland, cancer of the parathyroid gland,cancer of the adrenal gland, sarcoma of soft tissue, cancer of theurethra, cancer of the penis, chronic or acute leukemia, acute myeloidleukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL)(including non T cell ALL), chronic lymphocytic leukemia (CLL), solidtumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancerof the kidney or ureter, carcinoma of the renal pelvis, neoplasm of thecentral nervous system (CNS), primary CNS lymphoma, tumor angiogenesis,spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi'ssarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma,environmentally induced cancers including those induced by asbestos,other B cell malignancies, and combinations of said cancers. In oneparticular embodiment, the cancer is multiple myeloma. The particularcancer can be responsive to chemo- or radiation therapy or the cancercan be refractory. A refractory cancer refers to a cancer that is notamendable to surgical intervention and the cancer is either initiallyunresponsive to chemo- or radiation therapy or the cancer becomesunresponsive over time.

An “anti-tumor effect” as used herein, refers to a biological effectthat can present as a decrease in tumor volume, a decrease in the numberof tumor cells, a decrease in tumor cell proliferation, a decrease inthe number of metastases, an increase in overall or progression-freesurvival, an increase in life expectancy, or amelioration of variousphysiological symptoms associated with the tumor. An anti-tumor effectcan also refer to the prevention of the occurrence of a tumor, e.g., avaccine.

A “cytokine,” as used herein, refers to a non-antibody protein that isreleased by one cell in response to contact with a specific antigen,wherein the cytokine interacts with a second cell to mediate a responsein the second cell. A cytokine can be endogenously expressed by a cellor administered to a subject. Cytokines may be released by immune cells,including macrophages, B cells, T cells, and mast cells to propagate animmune response. Cytokines can induce various responses in the recipientcell. Cytokines can include homeostatic cytokines, chemokines,pro-inflammatory cytokines, effectors, and acute-phase proteins. Forexample, homeostatic cytokines, including interleukin (IL) 7 and IL-15,promote immune cell survival and proliferation, and pro-inflammatorycytokines can promote an inflammatory response. Examples of homeostaticcytokines include, but are not limited to, IL-2, IL-4, IL-5, IL-7,IL-10, IL-12p40, IL-12p70, IL-15, and interferon (IFN) gamma. Examplesof pro-inflammatory cytokines include, but are not limited to, IL-1a,IL-1b, IL-6, IL-13, IL-17a, tumor necrosis factor (TNF)-alpha, TNF-beta,fibroblast growth factor (FGF) 2, granulocyte macrophagecolony-stimulating factor (GM-CSF), soluble intercellular adhesionmolecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1),vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D, and placentalgrowth factor (PLGF). Examples of effectors include, but are not limitedto, granzyme A, granzyme B, soluble Fas ligand (sFasL), and perforin.Examples of acute phase-proteins include, but are not limited to,C-reactive protein (CRP) and serum amyloid A (SAA).

“Chemokines” are a type of cytokine that mediates cell chemotaxis, ordirectional movement. Examples of chemokines include, but are notlimited to, IL-8, IL-16, eotaxin, eotaxin-3, macrophage-derivedchemokine (MDC or CCL22), monocyte chemotactic protein 1 (MCP-1 orCCL2), MCP-4, macrophage inflammatory protein 1α (MIP-1α, MIP-1a),MIP-1β (MIP-1b), gamma-induced protein 10 (IP-10), and thymus andactivation regulated chemokine (TARC or CCL17).

A “therapeutically effective amount,” “effective dose,” “effectiveamount,” or “therapeutically effective dosage” of a therapeutic agent,e.g., engineered CAR T cells, is any amount that, when used alone or incombination with another therapeutic agent, protects a subject againstthe onset of a disease or promotes disease regression evidenced by adecrease in severity of disease symptoms, an increase in frequency andduration of disease symptom-free periods, or a prevention of impairmentor disability due to the disease affliction. The ability of atherapeutic agent to promote disease regression can be evaluated using avariety of methods known to the skilled practitioner, such as in humansubjects during clinical trials, in animal model systems predictive ofefficacy in humans, or by assaying the activity of the agent in in vitroassays.

The term “lymphocyte” as used herein includes natural killer (NK) cells,T cells, or B cells. NK cells are a type of cytotoxic (cell toxic)lymphocyte that represent a major component of the inherent immunesystem. NK cells reject tumors and cells infected by viruses. It worksthrough the process of apoptosis or programmed cell death. They weretermed “natural killers” because they do not require activation in orderto kill cells. T-cells play a major role in cell-mediated-immunity (noantibody involvement). Its T-cell receptors (TCR) differentiatethemselves from other lymphocyte types. The thymus, a specialized organof the immune system, is primarily responsible for the T cell'smaturation. There are six types of T-cells, namely: Helper T-cells(e.g., CD4+ cells), Cytotoxic T-cells (also known as TC, cytotoxic Tlymphocyte, CTL, T-killer cell, cytolytic T cell, CD8+ T-cells or killerT cell), Memory T-cells ((i) stem memory T_(SCM) cells, like naivecells, are CD45RO−, CCR7+, CD45RA+, CD62L+ (L-selectin), CD27+, CD28+and IL-7Rα+, but they also express large amounts of CD95, IL-2Rβ, CXCR3,and LFA-1, and show numerous functional attributes distinctive of memorycells); (ii) central memory T_(CM) cells express L-selectin and theCCR7, they secrete IL-2, but not IFNγ or IL-4, and (iii) effector memoryTEM cells, however, do not express L-selectin or CCR7 but produceeffector cytokines like IFNγ and IL-4), Regulatory T-cells (Tregs,suppressor T cells, or CD4+CD25+ regulatory T cells), Natural KillerT-cells (NKT) and Gamma Delta T-cells. B-cells, on the other hand, playa principal role in humoral immunity (with antibody involvement). Itmakes antibodies and antigens and performs the role ofantigen-presenting cells (APCs) and turns into memory B-cells afteractivation by antigen interaction. In mammals, immature B-cells areformed in the bone marrow, where its name is derived from.

The term “genetically engineered” or “engineered” refers to a method ofmodifying the genome of a cell, including, but not limited to, deletinga coding or non-coding region or a portion thereof or inserting a codingregion or a portion thereof. In some embodiments, the cell that ismodified is a lymphocyte, e.g., a T cell, which can either be obtainedfrom a patient or a donor. The cell can be modified to express anexogenous construct, such as, e.g., a chimeric antigen receptor (CAR) ora T cell receptor (TCR), which is incorporated into the cell's genome.

An “immune response” refers to the action of a cell of the immune system(for example, T lymphocytes, B lymphocytes, natural killer (NK) cells,macrophages, eosinophils, mast cells, dendritic cells and neutrophils)and soluble macromolecules produced by any of these cells or the liver(including Abs, cytokines, and complement) that results in selectivetargeting, binding to, damage to, destruction of, and/or eliminationfrom a vertebrate's body of invading pathogens, cells or tissuesinfected with pathogens, cancerous or other abnormal cells, or, in casesof autoimmunity or pathological inflammation, normal human cells ortissues.

The term “immunotherapy” refers to the treatment of a subject afflictedwith, or at risk of contracting or suffering a recurrence of, a diseaseby a method comprising inducing, enhancing, suppressing or otherwisemodifying an immune response. Examples of immunotherapy include, but arenot limited to, T cell therapies. T cell therapy can include adoptive Tcell therapy, tumor-infiltrating lymphocyte (TIL) immunotherapy,autologous cell therapy, engineered autologous cell therapy (eACT™), andallogeneic T cell transplantation. However, one of skill in the artwould recognize that the conditioning methods disclosed herein wouldenhance the effectiveness of any transplanted T cell therapy. Examplesof T cell therapies are described in U.S. Patent Publication Nos.2014/0154228 and 2002/0006409, U.S. Pat. No. 5,728,388, andInternational Publication No. WO 2008/081035.

The T cells of the immunotherapy can come from any source known in theart. For example, T cells can be differentiated in vitro from ahematopoietic stem cell population, or T cells can be obtained from asubject. T cells can be obtained from, e.g., peripheral bloodmononuclear cells (PBMCs), bone marrow, lymph node tissue, cord blood,thymus tissue, tissue from a site of infection, ascites, pleuraleffusion, spleen tissue, and tumors. In addition, the T cells can bederived from one or more T cell lines available in the art. T cells canalso be obtained from a unit of blood collected from a subject using anynumber of techniques known to the skilled artisan, such as FICOLL™separation and/or apheresis. Additional methods of isolating T cells fora T cell therapy are disclosed in U.S. Patent Publication No.2013/0287748, which is herein incorporated by references in itsentirety.

The term “engineered Autologous Cell Therapy,” which can be abbreviatedas “eACT™,” also known as adoptive cell transfer, is a process by whicha patient's own T cells are collected and subsequently geneticallyaltered to recognize and target one or more antigens expressed on thecell surface of one or more specific tumor cells or malignancies. Tcells can be engineered to express, for example, chimeric antigenreceptors (CAR) or T cell receptor (TCR). CAR positive (+) T cells areengineered to express an extracellular single chain variable fragment(scFv) with specificity for a particular tumor antigen linked to anintracellular signaling part comprising at least one costimulatorydomain and at least one activating domain. The costimulatory domain canbe derived from a naturally-occurring costimulatory domain, e.g., havingthe amino acid sequence of SEQ ID NO: 1, or a variant thereof, e.g., avariant having a truncated hinge domain (“THD”), and the activatingdomain can be derived from, e.g., CD3-zeta. In certain embodiments, theCAR is designed to have two, three, four, or more costimulatory domains.The CAR scFv can be designed to target, for example, CD19, which is atransmembrane protein expressed by cells in the B cell lineage,including all normal B cells and B cell malignances, including but notlimited to NHL, CLL, and non-T cell ALL. In some embodiments, the CAR isengineered such that the costimulatory domain is expressed as a separatepolypeptide chain. Example CART cell therapies and constructs aredescribed in U.S. Patent Publication Nos. 2013/0287748, 2014/0227237,2014/0099309, and 2014/0050708, and these references are incorporated byreference in their entirety.

A “patient” as used herein includes any human who is afflicted with acancer (e.g., a lymphoma or a leukemia). The terms “subject” and“patient” are used interchangeably herein.

As used herein, the term “in vitro cell” refers to any cell which iscultured ex vivo. In particular, an in vitro cell can include a T cell.

The terms “peptide,” “polypeptide,” and “protein” are usedinterchangeably, and refer to a compound comprised of amino acidresidues covalently linked by peptide bonds. A protein or peptidecontains at least two amino acids, and no limitation is placed on themaximum number of amino acids that can comprise a protein's or peptide'ssequence. Polypeptides include any peptide or protein comprising two ormore amino acids joined to each other by peptide bonds. As used herein,the term refers to both short chains, which also commonly are referredto in the art as peptides, oligopeptides and oligomers, for example, andto longer chains, which generally are referred to in the art asproteins, of which there are many types. “Polypeptides” include, forexample, biologically active fragments, substantially homologouspolypeptides, oligopeptides, homodimers, heterodimers, variants ofpolypeptides, modified polypeptides, derivatives, analogs, fusionproteins, among others. The polypeptides include natural peptides,recombinant peptides, synthetic peptides, or a combination thereof.

“Stimulation,” as used herein, refers to a primary response induced bybinding of a stimulatory molecule with its cognate ligand, wherein thebinding mediates a signal transduction event. A “stimulatory molecule”is a molecule on a T cell, e.g., the T cell receptor (TCR)/CD3 complex,that specifically binds with a cognate stimulatory ligand present on anantigen present cell. A “stimulatory ligand” is a ligand that whenpresent on an antigen presenting cell (e.g., an APC, a dendritic cell, aB-cell, and the like) can specifically bind with a stimulatory moleculeon a T cell, thereby mediating a primary response by the T cell,including, but not limited to, activation, initiation of an immuneresponse, proliferation, and the like. Stimulatory ligands include, butare not limited to, an anti-CD3 antibody (such as OKT3), an MHC Class Imolecule loaded with a peptide, a superagonist anti-CD2 antibody, and asuperagonist anti-CD28 antibody.

A “costimulatory signal,” as used herein, refers to a signal, which incombination with a primary signal, such as TCR/CD3 ligation, leads to aT cell response, such as, but not limited to, proliferation and/orupregulation or down regulation of key molecules.

A “costimulatory ligand” as used herein, includes a molecule on anantigen presenting cell that specifically binds a cognate co-stimulatorymolecule on a T cell. Binding of the costimulatory ligand provides asignal that mediates a T cell response, including, but not limited to,proliferation, activation, differentiation, and the like. Acostimulatory ligand induces a signal that is in addition to the primarysignal provided by a stimulatory molecule, for instance, by binding of aT cell receptor (TCR)/CD3 complex with a major histocompatibilitycomplex (MHC) molecule loaded with peptide. A co-stimulatory ligand caninclude, but is not limited to, 3/TR6, 4-1BB ligand, agonist or antibodythat binds Toll ligand receptor, B7-1 (CD80), B7-2 (CD86), CD30 ligand,CD40, CD7, CD70, CD83, herpes virus entry mediator (HVEM), humanleukocyte antigen G (HLA-G), ILT4, immunoglobulin-like transcript (ILT)3, inducible costimulatory ligand (ICOS-L), intercellular adhesionmolecule (ICAM), ligand that specifically binds with B7-H3, lymphotoxinbeta receptor, MHC class I chain-related protein A (MICA), MHC class Ichain-related protein B (MICB), OX40 ligand, PD-L2, or programmed death(PD) L1. A co-stimulatory ligand includes, without limitation, anantibody that specifically binds with a co-stimulatory molecule presenton a T cell, such as, but not limited to, 4-1BB, B7-H3, CD2, CD27, CD28,CD30, CD40, CD7, ICOS, ligand that specifically binds with CD83,lymphocyte function-associated antigen-1 (LFA-1), natural killer cellreceptor C (NKG2C), OX40, PD-1, or tumor necrosis factor superfamilymember 14 (TNFSF14 or LIGHT).

A “costimulatory molecule” is a cognate binding partner on a T cell thatspecifically binds with a costimulatory ligand, thereby mediating acostimulatory response by the T cell, such as, but not limited to,proliferation. Costimulatory molecules include, but are not limited to,A “costimulatory molecule” is a cognate binding partner on a T cell thatspecifically binds with a costimulatory ligand, thereby mediating acostimulatory response by the T cell, such as, but not limited to,proliferation. Costimulatory molecules include, but are not limited to,4-1BB/CD137, B7-H3, BAFFR, BLAME (SLAMF8), BTLA, CD 33, CD 45, CD100(SEMA4D), CD103, CD134, CD137, CD154, CD16, CD160 (BY55), CD18, CD19,CD19a, CD2, CD22, CD247, CD27, CD276 (B7-H3), CD28, CD29, CD3 (alpha;beta; delta; epsilon; gamma; zeta), CD30, CD37, CD4, CD4, CD40, CD49a,CD49D, CD49f, CD5, CD64, CD69, CD7, CD80, CD83 ligand, CD84, CD86,CD8alpha, CD8beta, CD9, CD96 (Tactile), CD1-1a, CD1-1b, CD1-1c, CD1-1d,CDS, CEACAM1, CRT AM, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS,GITR, HVEM (LIGHTR), IA4, ICAM-1, ICAM-1, ICOS, Ig alpha (CD79a), IL2Rbeta, IL2R gamma, IL7R alpha, integrin, ITGA4, ITGA4, ITGA6, ITGAD,ITGAE, ITGAL, ITGAM, ITGAX, ITGB2, ITGB7, ITGB1, KIRDS2, LAT, LFA-1,LFA-1, LIGHT, LIGHT (tumor necrosis factor superfamily member 14;TNFSF14), LTBR, Ly9 (CD229), lymphocyte function-associated antigen-1(LFA-1 (CD11a/CD18), MHC class I molecule, NKG2C, NKG2D, NKp30, NKp44,NKp46, NKp80 (KLRF1), OX40, PAG/Cbp, PD-1, PSGL1, SELPLG (CD162),signaling lymphocytic activation molecule, SLAM (SLAMF1; CD150; IPO-3),SLAMF4 (CD244; 2B4), SLAMF6 (NTB-A; Ly108), SLAMF7, SLP-76, TNF, TNFr,TNFR2, Toll ligand receptor, TRANCE/RANKL, VLA1, or VLA-6, or fragments,truncations, or combinations thereof.

The terms “reducing” and “decreasing” are used interchangeably hereinand indicate any change that is less than the original. “Reducing” and“decreasing” are relative terms, requiring a comparison between pre- andpost-measurements. “Reducing” and “decreasing” include completedepletions.

“Treatment” or “treating” of a subject refers to any type ofintervention or process performed on, or the administration of an activeagent to, the subject with the objective of reversing, alleviating,ameliorating, inhibiting, slowing down or preventing the onset,progression, development, severity or recurrence of a symptom,complication or condition, or biochemical indicia associated with adisease. In one embodiment, “treatment” or “treating” includes a partialremission. In another embodiment, “treatment” or “treating” includes acomplete remission.

To calculate percent identity, the sequences being compared aretypically aligned in a way that gives the largest match between thesequences. One example of a computer program that can be used todetermine percent identity is the GCG program package, which includesGAP (Devereux et al., 1984, Nucl. Acid Res. 12:387; Genetics ComputerGroup, University of Wisconsin, Madison, Wis.). The computer algorithmGAP is used to align the two polypeptides or polynucleotides for whichthe percent sequence identity is to be determined. The sequences arealigned for optimal matching of their respective amino acid ornucleotide (the “matched span,” as determined by the algorithm). Incertain embodiments, a standard comparison matrix (see, Dayhoff et al.,1978, Atlas of Protein Sequence and Structure 5:345-352 for the PAM 250comparison matrix; Henikoff et al., 1992, Proc. Natl. Acad. Sci. U.S.A.89:10915-10919 for the BLOSUM 62 comparison matrix) is also used by thealgorithm.

Various aspects of the invention are described in further detail in thefollowing subsections.

I. Chimeric Antigen Receptors and T Cell Receptors

Chimeric antigen receptors (CARs or CAR-Ts) and T cell receptors (TCRs)are genetically engineered receptors. These engineered receptors can bereadily inserted into and expressed by immune cells, including T cellsin accordance with techniques known in the art. With a CAR, a singlereceptor can be programmed to both recognize a specific antigen and,when bound to that antigen, activate the immune cell to attack anddestroy the cell bearing that antigen. When these antigens exist ontumor cells, an immune cell that expresses the CAR can target and killthe tumor cell.

One aspect of the present invention is directed to polynucleotidesencoding chimeric antigen receptors (CARs) or T cell receptors (TCRs)comprising a costimulatory domain comprising a novel extracellulardomain comprising a truncated hinge domain (“THD”), and engineered Tcells comprising a costimulatory domain comprising the novel THD. Thecostimulatory domain can further comprise a transmembrane domain and/oran intracellular domain. In some embodiments, a CAR or TCR encoded bythe polynucleotide of the present invention further comprises an antigenbinding molecule that specifically binds to a target antigen. In someembodiments, the CAR or TCR encoded by the polynucleotide furthercomprises an activating domain. In one particular embodiment, the CAR orTCR encoded by the polynucleotide comprises (i) an antigen bindingmolecule that specifically binds to a target antigen, (ii) acostimulatory domain comprising an extracellular domain, a transmembranedomain, and an intracellular domain, and (iii) an activating domain,wherein the extracellular domain comprises, consists essentially of, orconsists of a THD described herein, e.g., SEQ ID NO: 3.

In some embodiments, an orientation of the CARs in accordance with theinvention comprises an antigen binding domain (such as scFv) in tandemwith a costimulatory domain and an activating domain. The costimulatorydomain can comprise one or more of an extracellular portion, atransmembrane portion, and an intracellular portion. In otherembodiments, multiple costimulatory domains can be utilized in tandem.

I.A. Costimulatory Domain.

Chimeric antigen receptors incorporates costimulatory (signaling)domains to increase their potency. See U.S. Pat. Nos. 7,741,465, and6,319,494, as well as Krause et al. and Finney et al. (supra), Song etal., Blood 119:696-706 (2012); Kalos et al., Sci Transl. Med. 3:95(2011); Porter et al., N. Engl. J. Med. 365:725-33 (2011), and Gross etal., Annu. Rev. Pharmacol. Toxicol. 56:59-83 (2016). The costimulatoryprotein having the amino acid sequence of SEQ ID NO: 1 is acostimulatory protein found naturally on T-cells. The complete nativeamino acid sequence of this costimulatory protein is described in NCBIReference Sequence: NP_006130.1. See FIG. 1A. The complete nativenucleic acid sequence of this costimulatory protein is described in NCBIReference Sequence: NM_006139.1.

Novel Extracellular Domain:

The present disclosure shows that a novel extracellular domain of acostimulatory protein and comprising a truncated hinge domain (“THD”)can improve one or more properties of a CAR or a TCR. In someembodiments, the THD domain is a truncated version of a complete hingedomain (“CHD”). In certain embodiments, the isolated polynucleotideencoding a THD comprises (i) an amino acid sequence at least about 80%,at least about 85%, at least about 90%, at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or about 100% identical to amino acids 123 to 152 of SEQ ID NO: 1,wherein the THD domain does not contain amino acids 1 to 122 of SEQ IDNO: 1.

In other embodiments, the THD consists essentially of or consists of anamino acid sequence at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 96%, at least about 97%,at least about 98%, at least about 99%, or about 100% identical to aminoacids 123 to 152 of SEQ ID NO: 1. In other embodiments, the THD consistsessentially of or consists of an amino acid sequence encoded by anucleotide sequence at least about 60%, at least about 70%, at leastabout 80%, at least about 85%, at least about 90%, at least about 95%,at least about 96%, at least about 97%, at least about 98%, at leastabout 99%, or about 100% identical to SEQ ID NO: 3.

In some embodiments, the isolated polynucleotide encoding a THD consistsessentially of or consists of (i) an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to amino acids 123 to 152 of SEQ ID NO: 1and (ii) optionally ±one amino acid, ±two amino acids, ±three aminoacids, ±four amino acids, ±five amino acids, or ±six amino acids. Insome embodiments, the isolated polynucleotide encoding a THD consistsessentially of or consists of (i) an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to amino acids 123 to 152 of SEQ ID NO: 1and (ii) optionally one or two amino acids, one to three amino acids,one to four amino acids, one to five amino acids, or one to six aminoacids. The one to six amino acids that can be added or deleted from theamino acid sequence in the THD can be at either the N-terminus, at theC-terminus, or both the N-terminus and the C-terminus.

In some embodiments, the isolated polynucleotide encoding a THD consistsessentially of or consists of (i) an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to amino acids 123 to 152 of SEQ ID NO: 1and (ii) one additional N-terminal amino acid, two additional N-terminalamino acids, three additional N-terminal amino acids, four additionalN-terminal amino acids, five additional N-terminal amino acids, or sixadditional N-terminal amino acids.

In some embodiments, the additional amino acids can be N-terminal aminoacids. In some embodiments, the additional amino acids can beheterologous. In other embodiments, the additional amino acids are partof the naturally occurring costimulatory protein sequence.

In some embodiments, the THD consists essentially of or consists of anamino acid sequence at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 96%, at least about 97%,at least about 98%, at least about 99%, or about 100% identical to aminoacids 123 to 152 of SEQ ID NO: 1, amino acids 122 to 152 of SEQ ID NO:1, amino acids 121 to 152 of SEQ ID NO: 1, amino acids 120 to 152 of SEQID NO: 1, amino acids 119 to 152 of SEQ ID NO: 1, amino acids 118 to 152of SEQ ID NO: 1, or amino acids 117 to 152 of SEQ ID NO: 1.

In other embodiments, the THD consists essentially of or consists of anamino acid sequence at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 96%, at least about 97%,at least about 98%, at least about 99%, or about 100% identical to aminoacids 124 to 152 of SEQ ID NO: 1, amino acids 125 to 152 of SEQ ID NO:1, amino acids 126 to 152 of SEQ ID NO: 1, amino acids 127 to 152 of SEQID NO: 1, amino acids 128 to 152 of SEQ ID NO: 1, amino acids 129 to 152of SEQ ID NO: 1, or amino acids 130 to 152 of SEQ ID NO: 1.

In some embodiments, the THD does not comprise amino acids 1-116 of SEQID NO: 1. In some embodiments, the THD does not comprise amino acids1-117 of SEQ ID NO: 1. In some embodiments, the THD does not compriseamino acids 1-118 of SEQ ID NO: 1. In some embodiments, the THD does notcomprise amino acids 1-119 of SEQ ID NO: 1. In some embodiments, the THDdoes not comprise amino acids 1-120 of SEQ ID NO: 1. In someembodiments, the THD does not comprise amino acids 1-121 of SEQ IDNO: 1. In some embodiments, the THD does not comprise amino acids 1-122of SEQ ID NO: 1. In some embodiments, the THD does not comprise aminoacids 1-123 of SEQ ID NO: 1. In some embodiments, the THD does notcomprise amino acids 1-124 of SEQ ID NO: 1. In some embodiments, the THDdoes not comprise amino acids 1-125 of SEQ ID NO: 1. In someembodiments, the THD does not comprise amino acids 1-126 of SEQ IDNO: 1. In some embodiments, the THD does not comprise amino acids 1-127of SEQ ID NO: 1. In some embodiments, the THD does not comprise aminoacids 1-128 of SEQ ID NO: 1. In some embodiments, the THD does notcomprise amino acids 1-129 of SEQ ID NO: 1.

The corresponding amino acid sequence of the THD is set forth in SEQ IDNO. 3 LDNEKSNGTI IHVKGKHLCP SPLFPGPSKP. A nucleotide sequence encodingthe extracellular portion of THD is set forth in SEQ ID NO. 2CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCA.

In certain embodiments, the polynucleotide encoding a costimulatorydomain in a CAR or TCR comprises a nucleotide sequence at least about60%, at least about 70%, at least about 80%, at least about 85%, atleast about 90%, at least about 95%, at least about 96%, at least about97%, at least about 98%, at least about 99%, or about 100% identical toSEQ ID NO: 3, wherein the nucleotide sequence encodes a THD and whereinthe CAR or TCR does not comprise amino acids 1 to 122 of SEQ ID NO: 1.

In one particular embodiment, the THD consists essentially of orconsists of an amino acid sequence at least about 70%, at least about75%, at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or about 100% identical to the amino acidsequence of SEQ ID NO: 3. In a specific embodiment, the polynucleotideencoding THD consists essentially of or consists of a nucleotidesequence at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 85%, at least about90%, at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or about 100% identical to thenucleotide sequence of SEQ ID NO: 2.

In some embodiments, the THD further comprises some or all of a memberof the immunoglobulin family such as IgG1, IgG2, IgG3, IgG4, IgA, IgD,IgE, IgM, or fragment thereof.

In some embodiments, the THD is derived from a human complete hingedomain (“CHD”), e.g., from the costimulatory protein having the aminoacid sequence of SEQ ID NO: 1. In other embodiments, the THD is derivedfrom a rodent, murine, or primate (e.g., non-human primate) CHD of acostimulatory protein. In some embodiments, the THD is derived from achimeric CHD of a costimulatory protein.

Transmembrane Domain:

The costimulatory domain for the CAR or TCR of the invention can furthercomprise a transmembrane domain and/or an intracellular signalingdomain. The transmembrane domain can be designed to be fused to theextracellular domain of the CAR. It can similarly be fused to theintracellular domain of the CAR. In one embodiment, the transmembranedomain that naturally is associated with one of the domains in a CAR isused. In some instances, the transmembrane domain can be selected ormodified by amino acid substitution to avoid binding of such domains tothe transmembrane domains of the same or different surface membraneproteins to minimize interactions with other members of the receptorcomplex. The transmembrane domain can be derived either from a naturalor from a synthetic source. Where the source is natural, the domain canbe derived from any membrane-bound or transmembrane protein.Transmembrane regions of particular use in this invention can be derivedfrom (i.e., comprise) 4-1BB/CD137, activating NK cell receptors, anImmunoglobulin protein, B7-H3, BAFFR, BLAME (SLAMF8), BTLA, CD100(SEMA4D), CD103, CD160 (BY55), CD18, CD19, CD19a, CD2, CD247, CD27,CD276 (B7-H3), CD28, CD29, CD3 delta, CD3 epsilon, CD3 gamma, CD30, CD4,CD40, CD49a, CD49D, CD49f, CD69, CD7, CD84, CD8alpha, CD8beta, CD96(Tactile), CD11a, CD11b, CD11c, CD11d, CDS, CEACAM1, CRT AM, cytokinereceptor, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS, GITR, HVEM(LIGHTR), IA4, ICAM-1, ICAM-1, Ig alpha (CD79a), IL-2R beta, IL-2Rgamma, IL-7R alpha, inducible T cell costimulator (ICOS), integrins,ITGA4, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX, ITGB2, ITGB7,ITGB1, KIRDS2, LAT, LFA-1, LFA-1, a ligand that specifically binds withCD83, LIGHT, LIGHT, LTBR, Ly9 (CD229), lymphocyte function-associatedantigen-1 (LFA-1; CD1-1a/CD18), MHC class 1 molecule, NKG2C, NKG2D,NKp30, NKp44, NKp46, NKp80 (KLRF1), OX-40, PAG/Cbp, programmed death-1(PD-1), PSGL1, SELPLG (CD162), Signaling Lymphocytic ActivationMolecules (SLAM proteins), SLAM (SLAMF1; CD150; IPO-3), SLAMF4 (CD244;2B4), SLAMF6 (NTB-A; Ly108), SLAMF7, SLP-76, TNF receptor proteins,TNFR2, TNFSF14, a Toll ligand receptor, TRANCE/RANKL, VLA1, or VLA-6, ora fragment, truncation, or a combination thereof.

Optionally, short linkers can form linkages between any or some of theextracellular, transmembrane, and intracellular domains of the CAR.

In one specific embodiment, the nucleotide sequence of the costimulatoryprotein's transmembrane domain is set forth in SEQ ID NO. 4:TTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTT

In one embodiment, the polynucleotide encoding a transmembrane domainwithin a costimulatory domain comprises a nucleotide sequence at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, or about 100% identical to the nucleotide sequenceof SEQ ID NO: 4.

The amino acid sequence of the costimulatory protein's transmembranedomain is set forth in SEQ ID NO. 5: FWVLVVVGGV LACYSLLVTV AFIIFWV.

In one particular embodiment, the transmembrane domain within acostimulatory domain comprises an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to the amino acid sequence of SEQ ID NO: 5.

In another embodiment, the transmembrane domain is derived from (i.e.,comprises) CD8. In one embodiment, the nucleotide sequence of the CD8extracellular domain and transmembrane domain is set forth in SEQ ID NO:238 GCTGCAGCATTGAGCAACTCAATAATGTATTTTAGTCACTTTGTACCAGTGTTCTTGCCGGCTAAGCCTACTACCACACCCGCTCCACGGCCACCTACCCCAGCTCCTACCATCGCTTCACAGCCTCTGTCCCTGCGCCCAGAGGCTTGCCGACCGGCCGCAGGGGGCGCTGTTCATACCAGAGGACTGGATTTCGCCTGCGATATCTATATCTGGGCACCCCTGGCCGGAACCTGCGGCGTACTCCTGCTGTCCCTGGTCATCACGCTCTATTGT AATCACAGGAAC.

In some embodiments, the polynucleotide encoding a transmembrane domainwithin a costimulatory domain comprises a nucleotide sequence at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, or about 100% identical to the nucleotide sequenceof the CD8 transmembrane domain.

The amino acid sequence of the CD8 extracellular domain andtransmembrane domain is set forth in SEQ ID NO. 239AAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRN.

In one particular embodiment, the transmembrane domain within acostimulatory domain comprises an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to the amino acid sequence of the CD8transmembrane domain.

Intracellular (Signaling) Domain:

The intracellular (signaling) domain of the engineered T cells of theinvention can provide signaling to an activating domain, which thenactivates at least one of the normal effector functions of the immunecell. Effector function of a T cell, for example, can be cytolyticactivity or helper activity including the secretion of cytokines.

In certain embodiments, suitable intracellular signaling domain include(i.e., comprise), but are not limited to 4-1BB/CD137, activating NK cellreceptors, an Immunoglobulin protein, B7-H3, BAFFR, BLAME (SLAMF8),BTLA, CD100 (SEMA4D), CD103, CD160 (BY55), CD18, CD19, CD19a, CD2,CD247, CD27, CD276 (B7-H3), CD28, CD29, CD3 delta, CD3 epsilon, CD3gamma, CD30, CD4, CD40, CD49a, CD49D, CD49f, CD69, CD7, CD84, CD8alpha,CD8beta, CD96 (Tactile), CD11a, CD11b, CD11c, CD11d, CDS, CEACAM1, CRTAM, cytokine receptor, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS,GITR, HVEM (LIGHTR), IA4, ICAM-1, ICAM-1, Ig alpha (CD79a), IL-2R beta,IL-2R gamma, IL-7R alpha, inducible T cell costimulator (ICOS),integrins, ITGA4, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX,ITGB2, ITGB7, ITGB1, KIRDS2, LAT, LFA-1, LFA-1, ligand that specificallybinds with CD83, LIGHT, LIGHT, LTBR, Ly9 (CD229), Ly108), lymphocytefunction-associated antigen-1 (LFA-1; CD1-1a/CD18), MHC class 1molecule, NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), OX-40,PAG/Cbp, programmed death-1 (PD-1), PSGL1, SELPLG (CD162), SignalingLymphocytic Activation Molecules (SLAM proteins), SLAM (SLAMF1; CD150;IPO-3), SLAMF4 (CD244; 2B4), SLAMF6 (NTB-A, SLAMF7, SLP-76, TNF receptorproteins, TNFR2, TNFSF14, a Toll ligand receptor, TRANCE/RANKL, VLA1, orVLA-6, or a fragment, truncation, or a combination thereof.

An example of a nucleotide sequence encoding the intracellular signalingdomain is set forth in SEQ ID NO. 6:AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCG CTGCCTATCGGAGC

In one embodiment, the polynucleotide encoding an intracellularsignaling domain within a costimulatory domain comprises a nucleotidesequence at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 85%, at least about90%, at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or about 100% identical to thenucleotide sequence of SEQ ID NO: 6.

An example of an intracellular signaling domain is set forth in SEQ IDNO. 7: RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS.

In one particular embodiment, the intracellular signaling domain withina costimulatory domain comprises an amino acid sequence at least about80%, at least about 85%, at least about 90%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100% identical to the amino acid sequence of SEQ ID NO: 7.

In some embodiments, the costimulatory domain comprises, consistsessentially of, or consists of the extracellular THD, and thecostimulatory proteins's transmembrane and intracellular domains. Forexample, a nucleotide sequence encoding a costimulatory domain is setforth in SEQ ID NO. 240:CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

In some embodiments, the polynucleotide encoding a costimulatory domaincomprises, consists essentially of, or consists of a nucleotide sequenceat least about 60%, at least about 65%, at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%,at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, at least about 99%, or about 100% identical to the nucleotidesequence of SEQ ID NO: 240, wherein the costimulatory domain does notcomprises amino acids 1 to 122 of SEQ ID NO: 1, amino acids 1 to 121 ofSEQ ID NO: 1, amino acids 1 to 120 of SEQ ID NO: 1, amino acids 1 to 119of SEQ ID NO: 1, amino acids 1 to 118 of SEQ ID NO: 1, or amino acids 1to 118 of SEQ ID NO: 1.

The corresponding amino acid sequence of the costimulatory domain is setforth in SEQ ID NO. 241:LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

In some embodiments, the costimulatory domain comprises, consistsessentially of, or consists of a nucleotide sequence at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 96%, at least about 97%,at least about 98%, at least about 99%, or about 100% identical to theamino acid sequence of SEQ ID NO: 241, wherein the costimulatory domaindoes not comprises amino acids 1 to 122 of SEQ ID NO: 1, amino acids 1to 121 of SEQ ID NO: 1, amino acids 1 to 120 of SEQ ID NO: 1, aminoacids 1 to 119 of SEQ ID NO: 1, amino acids 1 to 118 of SEQ ID NO: 1, oramino acids 1 to 118 of SEQ ID NO: 1.

I.B. Activating Domain.

CD3 is an element of the T cell receptor on native T cells, and has beenshown to be an important intracellular activating element in CARs. Inone embodiment, the CD3 is CD3 zeta, the nucleotide sequence of which isset forth in SEQ ID NO. 8:AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAG CCCTGCCACCTAGG

In some embodiments, the polynucleotide encoding an activating domaincomprises a nucleotide sequence at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, at least about 99%, or about100% identical to the nucleotide sequence of SEQ ID NO: 8.

The corresponding amino acid of intracellular CD3 zeta is set forth inSEQ ID NO. 9: RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP PR

In some embodiments, the activating domain comprises a nucleotidesequence at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90%, at least about 95%, at least about96%, at least about 97%, at least about 98%, at least about 99%, orabout 100% identical to the amino acid sequence of SEQ ID NO: 9.

In some embodiments, the activating domain comprises an amino acidsequence at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90%, at least about 95%, at least about96%, at least about 97%, at least about 98%, at least about 99%, orabout 100% identical to the amino acid sequence of:

(SEQ ID NO: 251) RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDT YDALHMQALPPR.

I.C. Antigen Binding Molecules

CARs can be engineered to bind to an antigen (such as a cell-surfaceantigen) by incorporating an antigen binding molecule that interactswith that targeted antigen. In some embodiments, the antigen bindingmolecule is an antibody fragment thereof, e.g., one or more single chainantibody fragment (“scFv”). An scFv is a single chain antibody fragmenthaving the variable regions of the heavy and light chains of an antibodylinked together. See U.S. Pat. Nos. 7,741,465, and 6,319,494 as well asEshhar et al., Cancer Immunol Immunotherapy (1997) 45: 131-136. An scFvretains the parent antibody's ability to specifically interact withtarget antigen. scFvs are useful in chimeric antigen receptors becausethey can be engineered to be expressed as part of a single chain alongwith the other CAR components. Id. See also Krause et al., J. Exp. Med.,Volume 188, No. 4, 1998 (619-626); Finney et al., Journal of Immunology,1998, 161: 2791-2797. It will be appreciated that the antigen bindingmolecule is typically contained within the extracellular portion of theCAR such that it is capable of recognizing and binding to the antigen ofinterest. Bispecific and multispecific CARs are contemplated within thescope of the invention, with specificity to more than one target ofinterest.

In some embodiments, the polynucleotide encodes a CAR or a TCRcomprising a THD of the present invention and an antigen bindingmolecule that specifically binds to a target antigen. In someembodiments, the target antigen is a tumor antigen. In some embodiments,the antigen is selected from a tumor-associated surface antigen, such as5T4, alphafetoprotein (AFP), B7-1 (CD80), B7-2 (CD86), BCMA, B-humanchorionic gonadotropin, CA-125, carcinoembryonic antigen (CEA),carcinoembryonic antigen (CEA), CD123, CD133, CD138, CD19, CD20, CD22,CD23, CD24, CD25, CD30, CD33, CD34, CD4, CD40, CD44, CD56, CD8, CLL-1,c-Met, CMV-specific antigen, CSPG4, CTLA-4, disialoganglioside GD2,ductal-epithelial mucine, EBV-specific antigen, EGFR variant III(EGFRvIII), ELF2M, endoglin, ephrin B2, epidermal growth factor receptor(EGFR), epithelial cell adhesion molecule (EpCAM), epithelial tumorantigen, ErbB2 (HER2/neu), fibroblast associated protein (fap), FLT3,folate binding protein, GD2, GD3, glioma-associated antigen,glycosphingolipids, gp36, HBV-specific antigen, HCV-specific antigen,HER1-HER2, HER2-HER3 in combination, HERV-K, high molecularweight-melanoma associated antigen (HMW-MAA), HIV-1 envelopeglycoprotein gp41, HPV-specific antigen, human telomerase reversetranscriptase, IGFI receptor, IGF-II, IL-11Ralpha, IL-13R-a2, InfluenzaVirus-specific antigen; CD38, insulin growth factor (IGF1)-1, intestinalcarboxyl esterase, kappa chain, LAGA-1a, lambda chain, LassaVirus-specific antigen, lectin-reactive AFP, lineage-specific or tissuespecific antigen such as CD3, MAGE, MAGE-A1, major histocompatibilitycomplex (MHC) molecule, major histocompatibility complex (MHC) moleculepresenting a tumor-specific peptide epitope, M-CSF, melanoma-associatedantigen, mesothelin, mesothelin, MN-CA IX, MUC-1, mut hsp70-2, mutatedp53, mutated p53, mutated ras, neutrophil elastase, NKG2D, Nkp30,NY-ESO-1, p53, PAP, prostase, prostase specific antigen (PSA),prostate-carcinoma tumor antigen-1 (PCTA-1), prostate-specific antigen,prostein, PSMA, RAGE-1, ROR1, RU1, RU2 (AS), surface adhesion molecule,surviving and telomerase, TAG-72, the extra domain A (EDA) and extradomain B (EDB) of fibronectin and the A1 domain of tenascin-C (TnC A1),thyroglobulin, tumor stromal antigens, vascular endothelial growthfactor receptor-2 (VEGFR2), virus-specific surface antigen such as anHIV-specific antigen (such as HIV gp120), as well as any derivate orvariant of these surface markers. In certain embodiments, the antigenbinding molecule specifically binds to BCMA. In other embodiments, theantigen binding molecule specifically binds to CLL-1. In otherembodiments, the antigen binding molecule specifically binds to FLT3.

In some embodiments, the antigen binding molecule specifically bindsBCMA. In certain embodiments, the antigen binding molecule comprises (a)a VH CDR1 comprising an amino acid sequence selected from SEQ ID NOs:13-20; (b) a VH CDR2 comprising an amino acid sequence selected from SEQID NOs: 21-28; (c) a VH CDR3 comprising an amino acid sequence selectedfrom SEQ ID NOs: 29-36; (d) a VL CDR1 comprising an amino acid sequenceselected from SEQ ID NOs: 37-44; (e) a VL CDR2 comprising an amino acidsequence selected from SEQ ID NOs: 45-52; and/or (f) a VL CDR3comprising an amino acid sequence selected from SEQ ID NOs: 53-60.

In one embodiment, the antigen binding molecule comprises (a) a VH CDR1comprising an amino acid of SEQ ID NO: 13; (b) a VH CDR2 comprising anamino acid sequence of SEQ ID NO: 21; (c) a VH CDR3 comprising an aminoacid sequence of SEQ ID NO: 29; (d) a VL CDR1 comprising an amino acidsequence of SEQ ID NO: 37; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 45; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 53.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 14; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 22; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 30; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 38; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 46; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 54.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 15; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 23; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 31; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 39; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 47; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 55.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 16; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 24; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 32; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 40; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 48; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 56.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 17; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 25; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 33; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 41; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 49; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 57.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 18; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 26; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 34; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 42; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 50; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 58.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 19; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 27; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 35; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 43; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 51; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 59.

In another embodiment, the antigen binding molecule comprises (a) a VHCDR1 comprising an amino acid of SEQ ID NO: 20; (b) a VH CDR2 comprisingan amino acid sequence of SEQ ID NO: 28; (c) a VH CDR3 comprising anamino acid sequence of SEQ ID NO: 36; (d) a VL CDR1 comprising an aminoacid sequence of SEQ ID NO: 44; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 52; and/or (f) a VL CDR3 comprising an amino acidsequence of SEQ ID NO: 60.

In certain embodiments, the antigen binding molecule comprises a VHcomprising an amino acid sequence selected from the group consisting ofSEQ ID NOs: 77-84 and a VL comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 85-92. In one embodiment, theantigen binding molecule comprises a VH comprising an amino acidsequence of SEQ ID NO: 77 and a VL comprising an amino acid sequence ofSEQ ID NO: 85. In another embodiment, the antigen binding moleculecomprises a VH comprising an amino acid sequence of SEQ ID NO: 78 and aVL comprising an amino acid sequence of SEQ ID NO: 86. In anotherembodiment, the antigen binding molecule comprises a VH comprising anamino acid sequence of SEQ ID NO: 79 and a VL comprising an amino acidsequence of SEQ ID NO: 87. In another embodiment, the antigen bindingmolecule comprises a VH comprising an amino acid sequence of SEQ ID NO:80 and a VL comprising an amino acid sequence of SEQ ID NO: 88. Inanother embodiment, the antigen binding molecule comprises a VHcomprising an amino acid sequence of SEQ ID NO: 81 and a VL comprisingan amino acid sequence of SEQ ID NO: 89. In another embodiment, theantigen binding molecule comprises a VH comprising an amino acidsequence of SEQ ID NO: 82 and a VL comprising an amino acid sequence ofSEQ ID NO: 90. In another embodiment, the antigen binding moleculecomprises a VH comprising an amino acid sequence of SEQ ID NO: 83 and aVL comprising an amino acid sequence of SEQ ID NO: 91. In anotherembodiment, the antigen binding molecule comprises a VH comprising anamino acid sequence of SEQ ID NO: 84 and a VL comprising an amino acidsequence of SEQ ID NO: 92.

In one particular embodiment, the polynucleotide of the presentinvention comprises a nucleotide sequence at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%,at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, at least about 99%, or about 100% identical to a nucleotidesequence selected form the group consisting of SEQ ID NOs: 61-68. Inanother embodiment, the polynucleotide of the present inventioncomprises a nucleotide sequence at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, or about 100% identical to a nucleotide sequenceselected form the group consisting of SEQ ID NOs: 69-76.

Other known anti-BCMA antibodies or antigen binding molecules thereofcan be used as antigen binding molecules of a CAR or TCR comprising aTHD of the present invention. Non-limiting examples of such BCMAantibodies or antigen binding molecule thereof include antibodies orantigen binding molecules described in WO2015158671A1, published Oct.22, 2015 and WO2016014565A2, published Jan. 28, 2016.

In some embodiments, the antigen binding molecule specifically bindsCLL-1. In certain embodiments, the antigen binding molecule comprises(a) a VH CDR1 comprising an amino acid sequence selected from SEQ IDNOs: 93-96; (b) a VH CDR2 comprising an amino acid sequence selectedfrom SEQ ID NOs: 97-100; (c) a VH CDR3 comprising an amino acid sequenceselected from SEQ ID NOs: 101-104; (d) a VL CDR1 comprising an aminoacid sequence selected from SEQ ID NOs: 105-108; (e) a VL CDR2comprising an amino acid sequence selected from SEQ ID NOs: 109-112;and/or (f) a VL CDR3 comprising an amino acid sequence selected from SEQID NOs: 113-116.

In one embodiment, the antigen binding molecule comprises (a) a VH CDR1comprising an amino acid of SEQ ID NO: 93; (b) a VH CDR2 comprising anamino acid sequence of SEQ ID NO: 97; (c) a VH CDR3 comprising an aminoacid sequence of SEQ ID NO: 101; (d) a VL CDR1 comprising an amino acidsequence of SEQ ID NO: 105; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 109; and/or (f) a VL CDR3 comprising an aminoacid sequence of SEQ ID NO: 113.

In one embodiment, the antigen binding molecule comprises (a) a VH CDR1comprising an amino acid of SEQ ID NO: 94; (b) a VH CDR2 comprising anamino acid sequence of SEQ ID NO: 98; (c) a VH CDR3 comprising an aminoacid sequence of SEQ ID NO: 102; (d) a VL CDR1 comprising an amino acidsequence of SEQ ID NO: 106; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 110; and/or (f) a VL CDR3 comprising an aminoacid sequence of SEQ ID NO: 114.

In one embodiment, the antigen binding molecule comprises (a) a VH CDR1comprising an amino acid of SEQ ID NO: 95; (b) a VH CDR2 comprising anamino acid sequence of SEQ ID NO: 99; (c) a VH CDR3 comprising an aminoacid sequence of SEQ ID NO: 103; (d) a VL CDR1 comprising an amino acidsequence of SEQ ID NO: 107; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 111; and/or (f) a VL CDR3 comprising an aminoacid sequence of SEQ ID NO: 115.

In one embodiment, the antigen binding molecule comprises (a) a VH CDR1comprising an amino acid of SEQ ID NO: 96; (b) a VH CDR2 comprising anamino acid sequence of SEQ ID NO: 100; (c) a VH CDR3 comprising an aminoacid sequence of SEQ ID NO: 104; (d) a VL CDR1 comprising an amino acidsequence of SEQ ID NO: 108; (e) a VL CDR2 comprising an amino acidsequence of SEQ ID NO: 112; and/or (f) a VL CDR3 comprising an aminoacid sequence of SEQ ID NO: 116.

In certain embodiments, the antigen binding molecule comprises a VHcomprising an amino acid sequence selected from the group consisting ofSEQ ID NOs: 125-128 and a VL comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 129-132. In one embodiment, theantigen binding molecule comprises a VH comprising an amino acidsequence of SEQ ID NO: 125 and a VL comprising an amino acid sequence ofSEQ ID NO: 129. In another embodiment, the antigen binding moleculecomprises a VH comprising an amino acid sequence of SEQ ID NO: 126 and aVL comprising an amino acid sequence of SEQ ID NO: 130. In anotherembodiment, the antigen binding molecule comprises a VH comprising anamino acid sequence of SEQ ID NO: 127 and a VL comprising an amino acidsequence of SEQ ID NO: 131. In another embodiment, the antigen bindingmolecule comprises a VH comprising an amino acid sequence of SEQ ID NO:128 and a VL comprising an amino acid sequence of SEQ ID NO: 132.

In one particular embodiment, the polynucleotide of the presentinvention comprises a nucleotide sequence at least about 70%, at leastabout 75%, at least about 80%, at least about 85%, at least about 90%,at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, at least about 99%, or about 100% identical to a nucleotidesequence selected form the group consisting of SEQ ID NOs: 117-120. Inanother embodiment, the polynucleotide of the present inventioncomprises a nucleotide sequence at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 96%, at least about 97%, at least about 98%,at least about 99%, or about 100% identical to a nucleotide sequenceselected form the group consisting of SEQ ID NOs: 121-124.

Other examples of anti-CLL-1 antibodies or antigen binding moleculesthereof include antibodies or antigen binding molecules described inWO2016014535, published Jan. 28, 2016, and US 2016/0051651 A1, publishedFeb. 25, 2016.

The antigen binding molecule encoded by the polynucleotide of thepresent invention can be single chained or double chained. In someembodiments, the antigen binding molecule is single chained. In certainembodiments, the antigen binding molecule is selected from the groupconsisting of an scFv, an Fab, an Fab′, an Fv, an F(ab′)2, a dAb, andany combination thereof. In one particular embodiment, the antigenbinding molecule comprises an scFv.

In certain embodiments, the antigen binding molecule comprises a singlechain, wherein the heavy chain variable region and the light chainvariable region are connected by a linker. In some embodiments, the VHis located at the N terminus of the linker and the VL is located at theC terminus of the linker. In other embodiments, the VL is located at theN terminus of the linker and the VH is located at the C terminus of thelinker. In some embodiments, the linker comprises at least about 5, atleast about 8, at least about 10, at least about 13, at least about 15,at least about 18, at least about 20, at least about 25, at least about30, at least about 35, at least about 40, at least about 45, at leastabout 50, at least about 60, at least about 70, at least about 80, atleast about 90, or at least about 100 amino acids. In some embodiments,the linker comprises at least about 18 amino acids. In certainembodiments, the linker comprises an amino acid sequence that is atleast about 75%, at least about 85%, at least about 85%, at least about90%, at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or 100% identical to the amino acidsequence GSTSGSGKPGSGEGSTKG (SEQ ID NO: 12) or the amino acid sequenceGGGGSGGGGSGGGGS (SEQ ID NO: 237). In one embodiment, the linker is aWhitlow linker. In certain embodiments, the binding molecule comprises asingle chain, wherein the heavy chain variable region and the lightchain variable region are connected by a linker, wherein the linkercomprises the amino acid sequence of SEQ ID NO: 12.

In some embodiments, the antigen binding molecule binds a target antigen(e.g., human BCMA, human FLT3, or human CLL-1) with a K_(D) of less than1×10⁻⁷ M, less than 1×10⁻⁸ M, less than 1×10⁻⁷ M, or less than 1×10⁻⁹M.In one particular embodiment, the antigen binding molecule binds atarget antigen (e.g., human BCMA, human FLT3, or human CLL-1) with aK_(D) of less than 1×10⁻⁷ M. In another embodiment, the antigen bindingmolecule binds a target antigen (e.g., human BCMA, human FLT3, or humanCLL-1) with a K_(D) of less than 1×10⁻⁸ M. In some embodiments, theantigen binding molecule binds a target antigen (e.g., human BCMA, humanFLT3, or human CLL-1) with a K_(D) of about 1×10⁻⁷ M, about 2×10⁻⁷ M,about 3×10⁻⁷ M, about 4×10⁻⁷ M, about 5×10⁻⁷ M, about 6×10⁻⁷ M, about7×10⁻⁷ M, about 8×10⁻⁷ M, about 9×10⁻⁷ M, about 1×10⁻⁷ M, about 2×10⁻⁷M, about 3×10⁻⁷ M, about 4×10⁻⁸ M, about 5×10⁻⁸ M, about 6×10⁻⁸M, about7×10⁻⁸ M, about 8×10⁻⁸ M, about 9×10⁻⁸ M, about 1×10⁻⁹M, about 2×10⁻⁹ M,about 3×10⁻⁹ M, about 4×10⁻⁹ M, about 5×10⁻⁹ M, about 6×10⁻⁹ M, about7×10⁻⁹ M, about 8×10⁻⁹M, about 9×10⁻⁹ M, about 1×10⁻¹⁰ M, or about5×10⁻¹⁰ M. In certain embodiments, the K_(D) is calculated as thequotient of k_(off)/k_(on), and the k_(on) and k_(off) are determinedusing a monovalent antibody, such as a Fab fragment, as measured by,e.g., BIAcore® surface plasmon resonance technology. In otherembodiments, the K_(D) is calculated as the quotient of k_(off)/k_(on),and the k_(on) and k_(off) are determined using a bivalent antibody,such as a Fab fragment, as measured by, e.g., BIAcore® surface plasmonresonance technology.

In some embodiments, the antigen binding molecule binds a target antigen(e.g., human BCMA, human FLT3, or human CLL-1) with an association rate(k_(on)) of less than 1×10⁻⁴ M⁻¹ s⁻¹, less than 2×10⁻⁴ M⁻¹ s⁻¹ less than3×10⁻⁴ M⁻¹ s⁻¹ less than 4×10⁻⁴ M⁻¹ s⁻¹ less than 5×10⁻⁴ M⁻¹ s⁻¹, lessthan 6×10⁻⁴ M⁻¹ s⁻¹, less than 7×10⁻⁴ M⁻¹ s⁻¹, less than 8×10⁻⁴ M⁻¹ s⁻¹,less than 9×10⁻⁴ M⁻¹ s⁻¹, less than 1×10⁻⁵ M⁻¹ s⁻¹, less than 2×10⁻⁵ M⁻¹s⁻¹, less than 3×10⁻⁵ M⁻¹ s⁻¹, less than 4×10⁻⁵ M⁻¹ s⁻¹ less than 5×10⁻⁵M⁻¹ s⁻¹ less than 6×10⁻⁵ M⁻¹ s⁻¹ less than 7×10⁻⁵ M⁻¹ s⁻¹, less than8×10⁻⁵ M⁻¹ s⁻¹, less than 9×10⁻⁵ M⁻¹ s⁻¹, less than 1×10⁻⁶ M⁻¹ s⁻¹, lessthan 2×10⁻⁶ M⁻¹ s⁻¹, less than 3×10⁻⁶ M⁻¹ s⁻¹, less than 4×10⁻⁶ M⁻¹ s⁻¹,less than 5×10⁻⁶ M⁻¹ s⁻¹, less than 6×10⁻⁶ M⁻¹ s⁻¹, less than 7×10⁻⁶ M⁻¹s⁻¹, less than 8×10⁻⁶ M⁻¹ s⁻¹, less than 9×10⁻⁶ M⁻¹ s⁻¹, or less than1×10⁻⁷M⁻¹ s⁻¹. In certain embodiments, the k_(on) is determined using amonovalent antibody, such as a Fab fragment, as measured by, e.g.,BIAcore® surface plasmon resonance technology. In other embodiments, thek_(on) is determined using a bivalent antibody as measured by, e.g.,BIAcore® surface plasmon resonance technology.

In some embodiments, the antigen binding molecule binds a target antigen(e.g., human BCMA, human FLT3, or human CLL-1) with an dissociation rate(k_(off)) of less than 1×10⁻² s⁻¹, less than 2×10⁻² s⁻¹, less than3×10⁻² s⁻¹, less than 4×10⁻² s⁻¹, less than 5×10⁻² s⁻¹, less than 6×10⁻²s⁻¹, less than 7×10⁻² s⁻¹, less than 8×10⁻² s⁻¹, less than 9×10⁻² s⁻¹,less than 1×10⁻³ s⁻¹, less than 2×10⁻³ s⁻¹, less than 3×10⁻³ s⁻¹, lessthan 4×10⁻³ s⁻¹, less than 5×10⁻³ s⁻¹, less than 6×10⁻³ s⁻¹, less than7×10⁻³ s⁻¹, less than 8×10⁻³ s⁻¹, less than 9×10⁻³ s⁻¹, less than 1×10⁻⁴s⁻¹, less than 2×10⁻⁴ s⁻¹, less than 3×10⁻⁴ s⁻¹, less than 4×10⁻⁴ s⁻¹,less than 5×10⁻⁴ s⁻¹, less than 6×10⁻⁴ s⁻¹, less than 7×10⁻⁴ s⁻¹, lessthan 8×10⁻⁴ s⁻¹, less than 9×10⁻⁴ s⁻¹, less than 1×10⁻⁴ s⁻¹, or lessthan 5×10⁻⁴ s⁻¹ In certain embodiments, the k_(off) is determined usinga monovalent antibody, such as a Fab fragment, as measured by, e.g.,BIAcore® surface plasmon resonance technology. In other embodiments, thek_(off) is determined using a bivalent antibody as measured by, e.g.,BIAcore® surface plasmon resonance technology.

In some embodiments, the polynucleotide encodes a TCR, wherein the TCRfurther comprises a fourth complementarity determining region (CDR4). Incertain embodiments, the polynucleotide encodes a TCR, wherein the TCRfurther comprises a constant region. In some embodiments, the constantregion is selected from a constant region of IgG1, IgG2, IgG3, IgG4,IgA, IgD, IgE, and IgM.

I.D. Switch Domain

It will be appreciated that adverse events may be minimized bytransducing the immune cells (containing one or more CARs or TCRs) witha suicide gene. It may also be desired to incorporate an inducible “on”or “accelerator” switch into the immune cells. Suitable techniquesinclude use of inducible caspase-9 (U.S. Appl. 2011/0286980) or athymidine kinase, before, after or at the same time, as the cells aretransduced with the CAR construct of the present invention. Additionalmethods for introducing suicide genes and/or “on” switches includeTALENS, zinc fingers, RNAi, siRNA, shRNA, antisense technology, andother techniques known in the art.

In accordance with the invention, additional on-off or other types ofcontrol switch techniques may be incorporated herein. These techniquesmay employ the use of dimerization domains and optional activators ofsuch domain dimerization. These techniques include, e.g., thosedescribed by Wu et al., Science 2014 350 (6258) utilizing FKBP/Rapalogdimerization systems in certain cells, the contents of which areincorporated by reference herein in their entirety. Additionaldimerization technology is described in, e.g., Fegan et al. Chem. Rev.2010, 110, 3315-3336 as well as U.S. Pat. Nos. 5,830,462; 5,834,266;5,869,337; and 6,165,787, the contents of which are also incorporated byreference herein in their entirety. Additional dimerization pairs mayinclude cyclosporine-A/cyclophilin, receptor, estrogen/estrogen receptor(optionally using tamoxifen), glucocorticoids/glucocorticoid receptor,tetracycline/tetracycline receptor, vitamin D/vitamin D receptor.Further examples of dimerization technology can be found in e.g., WO2014/127261, WO 2015/090229, US 2014/0286987, US 2015/0266973, US2016/0046700, U.S. Pat. No. 8,486,693, US 2014/0171649, and US2012/0130076, the contents of which are further incorporated byreference herein in their entirety.

I.E. Leader Peptide

In some embodiments, the polynucleotide of the present invention encodesa CAR or a TCR can further comprises a leader peptide (also referred toherein as a “signal peptide”). In certain embodiments, the leaderpeptide comprises an amino acid sequence that is at least about 75%, atleast about 80%, at least about 85%, at least about 90%, at least about95%, at least about 96%, at least about 97%, at least about 98%, atleast about 99%, or 100% identical to the amino acid sequenceMALPVTALLLPLALLLHAARP (SEQ ID NO: 11). In some embodiments, the leaderpeptide comprises the amino acid sequence of SEQ ID NO: 11.

In some embodiments, the polynucleotide of the present invention encodesa CAR or a TCR, wherein the CAR or the TCR comprises a leader peptide(P), an antigen binding molecule (B), a costimulatory protein'sextracellular domain (E), a transmembrane domain (T), a costimulatoryregion (C), and an activation domain (A), wherein the CAR is configuredaccording to the following: P-B-E-T-C-A. In some embodiments, theantigen binding molecule comprises a VH and a VL, wherein the CAR isconfigured according to the following: P-VH-VL-E-T-C-A orP-VL-VH-E-T-C-A. In some embodiments, the VH and the VL are connected bya linker (L), wherein the CAR is configured according to the following,from N-terminus to C-terminus: P-VH-L-VL-E-T-C-A or P-VH-L-VL-E-T-C-A.

In some embodiments, the polynucleotide of the present invention encodesa CAR, wherein the CAR comprises an amino acid sequence at least about75%, at least about 85%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or 100% identical to an amino acid sequenceselected from Table 2. In certain embodiments, the polynucleotide of thepresent invention encodes a CAR, wherein the CAR comprises an amino acidsequence selected from Table 2.

TABLE 2 Example CAR Sequences SEQ SEQ CAR ID Amino Acid ID ConstructNucleotide Sequence NO: Sequence NO: 10E3_CHDATGGCACTCCCCGTAACTGCTCTGCTGCT 242 MALPVTALLLPLALLL 243GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVTLKESGPVLGCCCGCAGGTGACCCTCAAAGAGTCTGGA VKPTETLTLTCTVSGFCCCGTGCTCGTAAAACCTACGGAGACCCT SLINARMGVSWIRQPPGACACTCACCTGCACAGTCTCCGGCTTCA GKALEWLAHIFSNAEKGCCTCATCAATGCCAGGATGGGAGTTTCC SYRTSLKSRLTISKDTTGGATCAGGCAACCGCCCGGAAAGGCCCT SKSQVVLTMTNMDPVDGGAATGGCTCGCACATATTTTCAGTAACG TATYYCARIPGYGGNGCTGAAAAAAGCTATCGGACTTCTCTGAAA DYHYYGMDVWGQGTTVAGTCGGCTCACGATTAGTAAGGACACATC TVSSGGGGSGGGGSGGCAAGAGCCAAGTGGTGCTTACGATGACTA GGSDIQMTQSPSSLSAACATGGACCCTGTGGATACTGCAACCTAT SLGDRVTITCRASQGITACTGTGCTCGAATCCCTGGTTATGGCGG RNDLGWYQQKPGKAPKAAATGGGGACTACCACTACTACGGTATGG RLIYASSTLQSGVPSRATGTCTGGGGCCAAGGGACCACGGTTACT FSGSGSGTEFTLTISSGTTTCAAGCGGAGGGGGAGGGAGTGGGGG LQPEDFATYYCLQHNNTGGCGGATCTGGCGGAGGAGGCAGCGATA FPWTFGQGTKVEIKRATCCAGATGACGCAGTCCCCTAGTTCACTT AAIEVMYPPPYLDNEKTCCGCATCCCTGGGGGATCGGGTTACCAT SNGTIIHVKGKHLCPSTACATGCCGCGCGTCACAGGGTATCCGGA PLFPGPSKPFWVLVVVATGATCTGGGATGGTACCAGCAGAAGCCG GGVLACYSLLVTVAFIGGAAAGGCTCCTAAGCGCCTCATCTACGC IFWVRSKRSRLLHSDYCAGCTCCACCCTGCAGAGTGGAGTGCCCT MNMTPRRPGPTRKHYQCCCGGTTTTCAGGCAGTGGCTCCGGTACG PYAPPRDFAAYRSRVKGAGTTTACTCTTACAATTAGCAGCCTGCA FSRSADAPAYQQGQNQGCCAGAAGATTTTGCAACTTACTACTGTT LYNELNLGRREEYDVLTGCAGCATAATAATTTCCCCTGGACCTTT DKRRGRDPEMGGKPRRGGTCAGGGCACCAAGGTGGAGATCAAAAG KNPQEGLYNELQKDKMAGCAGCCGCCATCGAAGTAATGTATCCCC AEAYSEIGMKGERRRGCCCCGTACCTTGACAATGAGAAGTCAAAT KGHDGLYQGLSTATKDGGAACCATTATCCATGTTAAGGGCAAACA TYDALHMQALPPRCCTCTGCCCTTCTCCACTGTTCCCTGGCC CTAGTAAGCCGTTTTGGGTGCTGGTGGTAGTCGGTGGGGTGCTGGCTTGTTACTCTCT TCTCGTGACCGTCGCCTTTATAATCTTTTGGGTCAGATCCAAAAGAAGCCGCCTGCTC CATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACC AGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCCGAGTGAAATTTTCTAG ATCAGCTGATGCTCCCGCCTATCAGCAGGGACAGAATCAACTTTACAATGAGCTGAAC CTGGGTCGCAGAGAAGAGTACGACGTTTTGGACAAACGCCGGGGCCGAGATCCTGAGA TGGGGGGGAAGCCGAGAAGGAAGAATCCTCAAGAAGGCCTGTACAACGAGCTTCAAAA AGACAAAATGGCTGAGGCGTACTCTGAGATCGGCATGAAGGGCGAGCGGAGACGAGGC AAGGGTCACGATGGCTTGTATCAGGGCCTGAGTACAGCCACAAAGGACACCTATGACG CCCTCCACATGCAGGCACTGCCCCCACGC TAG 10E3_THDATGGCACTCCCCGTAACTGCTCTGCTGCT 244 MALPVTALLLPLALLL 245GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVTLKESGPVLGCCCGCAAGTTACTTTGAAGGAGTCTGGA VKPTETLTLTCTVSGFCCTGTACTGGTGAAGCCAACCGAGACACT SLINARMGVSWIRQPPGACACTCACGTGTACAGTGAGTGGTTTTT GKALEWLAHIFSNAEKCCTTGATCAACGCAAGGATGGGCGTCAGC SYRTSLKSRLTISKDTTGGATCAGGCAACCCCCTGGCAAGGCTCT SKSQVVLTMTNMDPVDGGAATGGCTCGCTCACATATTCAGCAATG TATYYCARIPGYGGNGCCGAAAAAAGCTACCGGACAAGCCTGAAA DYHYYGMDVWGQGTTVTCCCGCCTGACTATTTCCAAGGACACTTC TVSSGGGGSGGGGSGGTAAGTCTCAGGTGGTGCTGACCATGACCA GGSDIQMTQSPSSLSAACATGGACCCGGTGGACACCGCCACCTAT SLGDRVTITCRASQGITACTGCGCAAGAATCCCTGGGTATGGTGG RNDLGWYQQKPGKAPKGAATGGTGACTACCATTATTATGGGATGG RLIYASSTLQSGVPSRATGTGTGGGGGCAAGGCACAACCGTAACG FSGSGSGTEFTLTISSGTCTCAAGCGGTGGGGGAGGCTCAGGGGG LQPEDFATYYCLQHNNCGGAGGCTCCGGAGGTGGCGGCTCCGACA FPWTFGQGTKVEIKRATTCAGATGACCCAAAGCCCGTCCAGCCTG AALDNEKSNGTIIHVKTCCGCCAGCCTGGGAGATAGAGTGACAAT GKHLCPSPLFPGPSKPCACGTGTAGAGCTTCCCAAGGGATAAGAA FWVLVVVGGVLACYSLATGATCTCGGGTGGTATCAGCAGAAGCCC LVTVAFIIFWVRSKRSGGCAAAGCCCCCAAAAGGCTTATATATGC RLLHSDYMNMTPRRPGTAGTAGTACACTGCAGTCTGGAGTTCCTT PTRKHYQPYAPPRDFACCCGATTTTCAGGTAGCGGCTCCGGTACA AYRSRVKFSRSADAPAGAGTTCACCCTCACGATAAGCTCACTCCA YQQGQNQLYNELNLGRGCCTGAGGATTTCGCAACGTACTACTGCC REEYDVLDKRRGRDPETCCAGCACAACAATTTTCCCTGGACTTTC MGGKPRRKNPQEGLYNGGCCAGGGCACCAAGGTGGAGATCAAGAG ELQKDKMAEAYSEIGMGGCCGCTGCCCTTGATAATGAAAAGTCAA KGERRRGKGHDGLYQGACGGAACAATCATTCACGTGAAGGGCAAG LSTATKDTYDALHMQACACCTCTGTCCGTCACCCTTGTTCCCTGG LPPR TCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCT CTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGC TCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTA CCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCCGAGTGAAATTTTCT AGATCAGCTGATGCTCCCGCCTATCAGCAGGGACAGAATCAACTTTACAATGAGCTGA ACCTGGGTCGCAGAGAAGAGTACGACGTTTTGGACAAACGCCGGGGCCGAGATCCTGA GATGGGGGGGAAGCCGAGAAGGAAGAATCCTCAAGAAGGCCTGTACAACGAGCTTCAA AAAGACAAAATGGCTGAGGCGTACTCTGAGATCGGCATGAAGGGCGAGCGGAGACGAG GCAAGGGTCACGATGGCTTGTATCAGGGCCTGAGTACAGCCACAAAGGACACCTATGA CGCCCTCCACATGCAGGCACTGCCCCCAC GCTAG8B5_CHD ATGGCACTCCCCGTAACTGCTCTGCTGCT 246 MALPVTALLLPLALLL 247GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQIQLVESGGGVGCCCGCAGATCCAGTTGGTGGAATCAGGG VQPGRSLRLSCVASGFGGCGGTGTGGTGCAGCCGGGTAGGAGCCT TFKNYGMHWVRQAPGKGAGACTGTCATGCGTGGCGTCTGGCTTCA GLEWVAVIWYDGSNEYCATTCAAGAACTACGGCATGCACTGGGTG YGDPVKGRFTISRDNSCGACAGGCCCCCGGAAAGGGTTTGGAGTG KNMLYLQMNSLRADDTGGTCGCCGTGATCTGGTACGACGGATCTA AVYYCARSGIAVAGAFATGAGTATTACGGAGATCCTGTGAAGGGA DYWGQGTLVTVSSGGGAGGTTCACCATCTCCCGCGACAATAGCAA GSGGGGSGGGGSEIVLAAATATGCTCTACCTGCAAATGAACTCAC TQSPDTLSLSPGEKATTCAGGGCGGATGATACGGCGGTCTACTAT LSCRASQSVSSSFLAWTGCGCTCGCTCAGGGATTGCTGTGGCCGG YQQKPGQAPSLLIYVACGCATTCGATTACTGGGGACAGGGTACCC SRRAAGIPDRFSGSGSTGGTGACAGTATCAAGCGGAGGCGGCGGC GTDFTLTISRLEPEDFTCTGGCGGCGGCGGATCTGGCGGGGGGGG GMFYCQHYGRTPFTFGAAGTGAGATTGTGTTGACACAGTCTCCCG PGTKVDIKRAAAIEVMATACCCTGTCACTGTCACCCGGCGAGAAG YPPPYLDNEKSNGTIIGCAACGCTGAGTTGCAGAGCAAGCCAGTC HVKGKHLCPSPLFPGPAGTCTCCTCTTCTTTTCTGGCCTGGTATC SKPFWVLVVVGGVLACAGCAAAAACCAGGTCAGGCACCATCTCTC YSLLVTVAFIIFWVRSCTGATTTACGTTGCCAGCAGACGGGCGGC KRSRLLHSDYMNMTPRTGGCATTCCCGACAGGTTCTCTGGAAGCG RPGPTRKHYQPYAPPRGATCTGGGACCGATTTTACCCTGACAATT DFAAYRSRVKFSRSADAGCCGCTTGGAGCCCGAAGACTTTGGTAT APAYQQGQNQLYNELNGTTTTACTGCCAGCACTACGGAAGGACAC LGRREEYDVLDKRRGRCTTTCACATTTGGCCCGGGCACGAAAGTC DPEMGGKPRRKNPQEGGATATAAAACGCGCAGCCGCCATTGAAGT LYNELQKDKMAEAYSEAATGTACCCACCACCTTATTTGGACAATG IGMKGERRRGKGHDGLAAAAGTCCAATGGTACCATTATTCACGTC YQGLSTATKDTYDALHAAGGGAAAGCATCTCTGTCCAAGCCCTCT MQALPPR GTTCCCCGGCCCCTCCAAACCATTCTGGGTGCTGGTGGTCGTCGGCGGAGTTCTGGCC TGCTATTCTCTGCTCGTGACTGTTGCATTCATCATTTTCTGGGTGAGATCCAAAAGAA GCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAG GAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCCGAGTG AAATTTTCTAGATCAGCTGATGCTCCCGCCTATCAGCAGGGACAGAATCAACTTTACA ATGAGCTGAACCTGGGTCGCAGAGAAGAGTACGACGTTTTGGACAAACGCCGGGGCCG AGATCCTGAGATGGGGGGGAAGCCGAGAAGGAAGAATCCTCAAGAAGGCCTGTACAAC GAGCTTCAAAAAGACAAAATGGCTGAGGCGTACTCTGAGATCGGCATGAAGGGCGAGC GGAGACGAGGCAAGGGTCACGATGGCTTGTATCAGGGCCTGAGTACAGCCACAAAGGA CACCTATGACGCCCTCCACATGCAGGCACTGCCCCCACGCTAG 8B5_THD ATGGCACTCCCCGTAACTGCTCTGCTGCT 248MALPVTALLLPLALLL 249 GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQIQLVESGGGVGCCCGCAGATTCAGCTCGTGGAGTCAGGT VQPGRSLRLSCVASGFGGTGGCGTGGTTCAGCCCGGACGGTCCCT TFKNYGMHWVRQAPGKGCGACTCTCTTGTGTGGCAAGCGGATTTA GLEWVAVIWYDGSNEYCCTTTAAGAACTATGGCATGCACTGGGTG YGDPVKGRFTISRDNSAGGCAGGCCCCTGGAAAAGGACTGGAGTG KNMLYLQMNSLRADDTGGTTGCTGTGATCTGGTACGACGGGTCCA AVYYCARSGIAVAGAFACGAATATTATGGCGATCCTGTGAAGGGA DYWGQGTLVTVSSGGGCGGTTTACAATCTCACGCGATAACTCAAA GSGGGGSGGGGSEIVLGAACATGCTGTACCTGCAAATGAACTCTC TQSPDTLSLSPGEKATTGCGCGCTGATGACACTGCCGTGTATTAT LSCRASQSVSSSFLAWTGCGCTCGGAGTGGTATCGCCGTCGCAGG YQQKPGQAPSLLIYVAAGCATTTGATTATTGGGGGCAAGGGACCC SRRAAGIPDRFSGSGSTCGTGACAGTGAGTTCCGGAGGGGGAGGT GTDFTLTISRLEPEDFTCTGGTGGAGGCGGCTCTGGTGGGGGAGG GMFYCQHYGRTPFTFGCAGCGAGATCGTTCTGACCCAGTCTCCTG PGTKVDIKRAAALDNEACACACTGTCACTGTCCCCTGGTGAAAAG KSNGTIIHVKGKHLCPGCCACACTGTCTTGTAGAGCGTCCCAGAG SPLFPGPSKPFWVLVVCGTTTCCAGTTCCTTCCTTGCATGGTATC VGGVLACYSLLVTVAFAACAAAAACCCGGGCAGGCTCCAAGCTTG IIFWVRSKRSRLLHSDCTGATCTACGTGGCCAGCCGCCGGGCCGC YMNMTPRRPGPTRKHYAGGCATCCCTGATAGGTTTAGCGGTTCTG QPYAPPRDFAAYRSRVGGAGCGGGACGGACTTCACCTTGACAATA KFSRSADAPAYQQGQNTCACGGCTGGAACCCGAAGACTTCGGAAT QLYNELNLGRREEYDVGTTTTATTGCCAGCACTACGGAAGAACTC LDKRRGRDPEMGGKPRCATTCACCTTTGGCCCGGGAACGAAGGTA RKNPQEGLYNELQKDKGACATCAAGAGAGCAGCAGCCCTCGACAA MAEAYSEIGMKGERRRCGAGAAATCCAATGGAACCATTATCCATG GKGHDGLYQGLSTATKTGAAGGGGAAACATCTCTGCCCTTCACCA DTYDALHMQALPPRTTGTTCCCTGGACCCAGCAAGCCTTTTTG GGTTCTGGTCGTGGTGGGGGGCGTCCTGGCTTGTTACTCCCTCCTCGTTACAGTCGCC TTCATAATCTTTTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGA ATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACC TAGAGATTTCGCTGCCTATCGGAGCCGAGTGAAATTTTCTAGATCAGCTGATGCTCCC GCCTATCAGCAGGGACAGAATCAACTTTACAATGAGCTGAACCTGGGTCGCAGAGAAG AGTACGACGTTTTGGACAAACGCCGGGGCCGAGATCCTGAGATGGGGGGGAAGCCGAG AAGGAAGAATCCTCAAGAAGGCCTGTACAACGAGCTTCAAAAAGACAAAATGGCTGAG GCGTACTCTGAGATCGGCATGAAGGGCGAGCGGAGACGAGGCAAGGGTCACGATGGCT TGTATCAGGGCCTGAGTACAGCCACAAAGGACACCTATGACGCCCTCCACATGCAGGC ACTGCCCCCACGCTAG FS-ATGGCACTCCCCGTAACTGCTCTGCTGCT 133 MALPVTALLLPLALLL 134 21495CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEVQLLESGGGL LGCCCGGAGGTGCAGCTGTTGGAGTCTGGG VQPGGSLRLSCAASGFGGAGGCTTGGTACAGCCTGGGGGGTCCCT TFSSYAMSWVRQAPGKGAGACTCTCCTGTGCAGCCTCTGGATTCA GLEWVSAISGSGGSTYCCTTTAGCAGCTATGCCATGAGCTGGGTC YADSVKGRFTISRDNSCGCCAGGCTCCAGGGAAGGGGCTGGAGTG KNTLYLQMNSLRAEDTGGTCTCAGCTATTAGTGGTAGTGGTGGTA AVYYCARAEMGAVFDIGCACATACTACGCAGACTCCGTGAAGGGC WGQGTMVTVSSGSTSGCGGTTCACCATCTCCAGAGACAATTCCAA SGKPGSGEGSTKGEIVGAACACGCTGTATCTGCAAATGAACAGCC LTQSPATLSLSPGERATGAGAGCCGAGGACACGGCGGTGTACTAC TLSCRASQSVSRYLAWTGCGCAAGAGCCGAGATGGGAGCCGTATT YQQKPGQAPRLLIYDACGACATATGGGGTCAGGGTACAATGGTCA SNRATGIPARFSGSGSCCGTCTCCTCAGGGTCTACATCCGGCTCC GTDFTLTISSLEPEDFGGGAAGCCCGGAAGTGGCGAAGGTAGTAC AVYYCQQRISWPFTFGAAAGGGGGAAATTGTGTTGACACAGTCTC GGTKVEIKRAAALDNECAGCCACCCTGTCTTTGTCTCCAGGGGAA KSNGTIIHVKGKHLCPAGAGCCACCCTCTCCTGCAGGGCCAGTCA SPLFPGPSKPFWVLVVGAGTGTTAGCAGGTACTTAGCCTGGTACC VGGVLACYSLLVTVAFAACAGAAACCTGGCCAGGCTCCCAGGCTC IIFWVRSKRSRLLHSDCTCATCTATGATGCATCCAACAGGGCCAC YMNMTPRRPGPTRKHYTGGCATCCCAGCCAGGTTCAGTGGCAGTG QPYAPPRDFAAYRSRVGGTCTGGGACAGACTTCACTCTCACCATC KFSRSADAPAYQQGQNAGCAGCCTAGAGCCTGAAGATTTTGCAGT QLYNELNLGRREEYDVTTATTACTGTCAGCAGAGAATCTCCTGGC LDKRRGRDPEMGGKPRCTTTCACTTTTGGCGGAGGGACCAAGGTT RKNPQEGLYNELQKDKGAGATCAAACGGGCCGCTGCCCTTGATAA MAEAYSEIGMKGERRRTGAAAAGTCAAACGGAACAATCATTCACG GKGHDGLYQGLSTATKTGAAGGGCAAGCACCTCTGTCCGTCACCC DTYDALHMQALPPRTTGTTCCCTGGTCCATCCAAGCCATTCTG GGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCT TTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGA ATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACC TAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCA GCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAG AGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAG ACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAA GCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTT TGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGC CCTGCCACCTAGGTAA FS-ATGGCACTCCCCGTAACTGCTCTGCTGCT 135 MALPVTALLLPLALLL 136 21495CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEIVLTQSPATL HGCCCGGAAATTGTGTTGACACAGTCTCCA SLSPGERATLSCRASQGCCACCCTGTCTTTGTCTCCAGGGGAAAG SVSRYLAWYQQKPGQAAGCCACCCTCTCCTGCAGGGCCAGTCAGA PRLLIYDASNRATGIPGTGTTAGCAGGTACTTAGCCTGGTACCAA ARFSGSGSGTDFTLTICAGAAACCTGGCCAGGCTCCCAGGCTCCT SSLEPEDFAVYYCQQRCATCTATGATGCATCCAACAGGGCCACTG ISWPFTFGGGTKVEIKGCATCCCAGCCAGGTTCAGTGGCAGTGGG RGSTSGSGKPGSGEGSTCTGGGACAGACTTCACTCTCACCATCAG TKGEVQLLESGGGLVQCAGCCTAGAGCCTGAAGATTTTGCAGTTT PGGSLRLSCAASGFTFATTACTGTCAGCAGAGAATCTCCTGGCCT SSYAMSWVRQAPGKGLTTCACTTTTGGCGGAGGGACCAAGGTTGA EWVSAISGSGGSTYYAGATCAAACGGGGGTCTACATCCGGCTCCG DSVKGRFTISRDNSKNGGAAGCCCGGAAGTGGCGAAGGTAGTACA TLYLQMNSLRAEDTAVAAGGGGGAGGTGCAGCTGTTGGAGTCTGG YYCARAEMGAVFDIWGGGGAGGCTTGGTACAGCCTGGGGGGTCCC QGTMVTVSSAAALDNETGAGACTCTCCTGTGCAGCCTCTGGATTC KSNGTIIHVKGKHLCPACCTTTAGCAGCTATGCCATGAGCTGGGT SPLFPGPSKPFWVLVVCCGCCAGGCTCCAGGGAAGGGGCTGGAGT VGGVLACYSLLVTVAFGGGTCTCAGCTATTAGTGGTAGTGGTGGT IIFWVRSKRSRLLHSDAGCACATACTACGCAGACTCCGTGAAGGG YMNMTPRRPGPTRKHYCCGGTTCACCATCTCCAGAGACAATTCCA QPYAPPRDFAAYRSRVAGAACACGCTGTATCTGCAAATGAACAGC KFSRSADAPAYQQGQNCTGAGAGCCGAGGACACGGCGGTGTACTA QLYNELNLGRREEYDVCTGCGCAAGAGCCGAGATGGGAGCCGTAT LDKRRGRDPEMGGKPRTCGACATATGGGGTCAGGGTACAATGGTC RKNPQEGLYNELQKDKACCGTCTCCTCAGCCGCTGCCCTTGATAA MAEAYSEIGMKGERRRTGAAAAGTCAAACGGAACAATCATTCACG GKGHDGLYQGLSTATKTGAAGGGCAAGCACCTCTGTCCGTCACCC DTYDALHMQALPPRTTGTTCCCTGGTCCATCCAAGCCATTCTG GGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCT TTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGA ATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACC TAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCA GCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAG AGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAG ACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAA GCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTT TGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGC CCTGCCACCTAGGTAA PC-ATGGCACTCCCCGTAACTGCTCTGCTGCT 137 MALPVTALLLPLALLL 138 21497CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVESGGGV LGCCCGCAGGTGCAGCTGGTGGAGTCTGGG VQPGRSLRLSCAASGFGGAGGCGTGGTCCAGCCTGGGAGGTCCCT TFSSYGMHWVRQAPGKGAGACTCTCCTGTGCAGCGTCTGGATTCA GLEWVAVISYDGSNKYCCTTCAGTAGCTATGGCATGCACTGGGTC YADSVKGRFTISRDNSCGCCAGGCTCCAGGCAAGGGGCTGGAGTG KNTLYLQMNSLRAEDTGGTGGCAGTTATATCGTATGATGGAAGTA AVYYCARDGTYLGGLWATAAATACTATGCAGACTCCGTGAAGGGC YFDLWGRGTLVTVSSGCGATTCACCATCTCCAGAGACAATTCCAA STSGSGKPGSGEGSTKGAACACGCTGTATCTGCAAATGAACAGCC GDIVMTQSPLSLPVTPTGAGAGCCGAGGACACGGCGGTGTACTAC GEPASISCRSSQSLLHTGCGCCAGAGACGGTACTTATCTAGGTGG SNGYNYLDWYLQKPGQTCTCTGGTACTTCGACTTATGGGGGAGAG SPQLLIYLGSNRASGVGTACCTTGGTCACCGTCTCCTCAGGGTCT PDRFSGSGSGTDFTLKACATCCGGCTCCGGGAAGCCCGGAAGTGG ISRVEAEDVGVYYCMQCGAAGGTAGTACAAAGGGGGATATTGTGA GLGLPLTFGGGTKVEITGACTCAGTCTCCACTCTCCCTGCCCGTC KRAAALDNEKSNGTIIACCCCTGGAGAGCCGGCCTCCATCTCCTG HVKGKHLCPSPLFPGPCAGGTCTAGTCAGAGCCTCCTGCATAGTA SKPFWVLVVVGGVLACATGGATACAACTATTTGGATTGGTACCTG YSLLVTVAFIIFWVRSCAGAAGCCAGGGCAGTCTCCACAGCTCCT KRSRLLHSDYMNMTPRGATCTATTTGGGTTCTAATCGGGCCTCCG RPGPTRKHYQPYAPPRGGGTCCCTGACAGGTTCAGTGGCAGTGGA DFAAYRSRVKFSRSADTCAGGCACAGATTTTACACTGAAAATCAG APAYQQGQNQLYNELNCAGAGTGGAGGCTGAGGATGTTGGGGTTT LGRREEYDVLDKRRGRATTACTGCATGCAGGGACTCGGCCTCCCT DPEMGGKPRRKNPQEGCTCACTTTTGGCGGAGGGACCAAGGTTGA LYNELQKDKMAEAYSEGATCAAACGGGCCGCTGCCCTTGATAATG IGMKGERRRGKGHDGLAAAAGTCAAACGGAACAATCATTCACGTG YQGLSTATKDTYDALHAAGGGCAAGCACCTCTGTCCGTCACCCTT MQALPPR GTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCT TGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAA GCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAG GAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTG AAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATA ACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACG GGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAAT GAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGC GGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGA TACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA PC- ATGGCACTCCCCGTAACTGCTCTGCTGCT 139 MALPVTALLLPLALLL140 21497CARHx GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPDIVMTQSPLSL LGCCCGGATATTGTGATGACTCAGTCTCCA PVTPGEPASISCRSSQCTCTCCCTGCCCGTCACCCCTGGAGAGCC SLLHSNGYNYLDWYLQGGCCTCCATCTCCTGCAGGTCTAGTCAGA KPGQSPQLLIYLGSNRGCCTCCTGCATAGTAATGGATACAACTAT ASGVPDRFSGSGSGTDTTGGATTGGTACCTGCAGAAGCCAGGGCA FTLKISRVEAEDVGVYGTCTCCACAGCTCCTGATCTATTTGGGTT YCMQGLGLPLTFGGGTCTAATCGGGCCTCCGGGGTCCCTGACAGG KVEIKRGSTSGSGKPGTTCAGTGGCAGTGGATCAGGCACAGATTT SGEGSTKGQVQLVESGTACACTGAAAATCAGCAGAGTGGAGGCTG GGVVQPGRSLRLSCAAAGGATGTTGGGGTTTATTACTGCATGCAG SGFTFSSYGMHWVRQAGGACTCGGCCTCCCTCTCACTTTTGGCGG PGKGLEWVAVISYDGSAGGGACCAAGGTTGAGATCAAACGGGGGT NKYYADSVKGRFTISRCTACATCCGGCTCCGGGAAGCCCGGAAGT DNSKNTLYLQMNSLRAGGCGAAGGTAGTACAAAGGGGCAGGTGCA EDTAVYYCARDGTYLGGCTGGTGGAGTCTGGGGGAGGCGTGGTCC GLWYFDLWGRGTLVTVAGCCTGGGAGGTCCCTGAGACTCTCCTGT SSAAALDNEKSNGTIIGCAGCGTCTGGATTCACCTTCAGTAGCTA HVKGKHLCPSPLFPGPTGGCATGCACTGGGTCCGCCAGGCTCCAG SKPFWVLVVVGGVLACGCAAGGGGCTGGAGTGGGTGGCAGTTATA YSLLVTVAFIIFWVRSTCGTATGATGGAAGTAATAAATACTATGC KRSRLLHSDYMNMTPRAGACTCCGTGAAGGGCCGATTCACCATCT RPGPTRKHYQPYAPPRCCAGAGACAATTCCAAGAACACGCTGTAT DFAAYRSRVKFSRSADCTGCAAATGAACAGCCTGAGAGCCGAGGA APAYQQGQNQLYNELNCACGGCGGTGTACTACTGCGCCAGAGACG LGRREEYDVLDKRRGRGTACTTATCTAGGTGGTCTCTGGTACTTC DPEMGGKPRRKNPQEGGACTTATGGGGGAGAGGTACCTTGGTCAC LYNELQKDKMAEAYSECGTCTCCTCAGCCGCTGCCCTTGATAATG IGMKGERRRGKGHDGLAAAAGTCAAACGGAACAATCATTCACGTG YQGLSTATKDTYDALHAAGGGCAAGCACCTCTGTCCGTCACCCTT MQALPPR GTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCT TGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAA GCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAG GAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTG AAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATA ACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACG GGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAAT GAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGC GGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGA TACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA AJ- ATGGCACTCCCCGTAACTGCTCTGCTGCT 141 MALPVTALLLPLALLL142 21508CARHx GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVQSGAEV LGCCCGCAGGTGCAGCTGGTGCAGTCTGGG KKPGASVKVSCKASGYGCTGAGGTGAAGAAGCCTGGGGCCTCAGT TFTSYYMHWVRQAPGQGAAGGTTTCCTGCAAGGCATCTGGATACA GLEWMGIINPGGGSTSCCTTCACCAGCTACTATATGCACTGGGTG YAQKFQGRVTMTRDTSCGACAGGCCCCTGGACAAGGGCTTGAGTG TSTVYMELSSLRSEDTGATGGGAATAATCAACCCTGGTGGTGGTA AVYYCARESWPMDVWGGCACAAGCTACGCACAGAAGTTCCAGGGC QGTTVTVSSGSTSGSGAGAGTCACCATGACCAGGGACACGTCCAC KPGSGEGSTKGEIVMTGAGCACAGTCTACATGGAGCTGAGCAGCC QSPATLSVSPGERATLTGAGATCTGAGGACACGGCGGTGTACTAC SCRASQSVSSNLAWYQTGCGCCAGAGAGAGTTGGCCAATGGACGT QKPGQAPRLLIYGASTATGGGGCCAGGGAACAACTGTCACCGTCT RATGIPARESGSGSGTCCTCAGGGTCTACATCCGGCTCCGGGAAG EFTLTISSLQSEDFAVCCCGGAAGTGGCGAAGGTAGTACAAAGGG YYCQQYAAYPTFGGGTGGAAATAGTGATGACGCAGTCTCCAGCCA KVEIKRAAALDNEKSNCCCTGTCTGTGTCTCCAGGGGAAAGAGCC GTIIHVKGKHLCPSPLACCCTCTCCTGCAGGGCCAGTCAGAGTGT FPGPSKPFWVLVVVGGTAGCAGCAACTTAGCCTGGTACCAGCAGA VLACYSLLVTVAFIIFAACCTGGCCAGGCTCCCAGGCTCCTCATC WVRSKRSRLLHSDYMNTATGGTGCATCCACCAGGGCCACTGGTAT MTPRRPGPTRKHYQPYCCCAGCCAGGTTCAGTGGCAGTGGGTCTG APPRDFAAYRSRVKFSGGACAGAGTTCACTCTCACCATCAGCAGC RSADAPAYQQGQNQLYCTGCAGTCTGAAGATTTTGCAGTTTATTA NELNLGRREEYDVLDKCTGTCAGCAGTACGCCGCCTACCCTACTT RRGRDPEMGGKPRRKNTTGGCGGAGGGACCAAGGTTGAGATCAAA PQEGLYNELQKDKMAECGGGCCGCTGCCCTTGATAATGAAAAGTC AYSEIGMKGERRRGKGAAACGGAACAATCATTCACGTGAAGGGCA HDGLYQGLSTATKDTYAGCACCTCTGTCCGTCACCCTTGTTCCCT DALHMQALPPR GGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACT CTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCT GCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACAC TACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTT CCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCT CAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCT GAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGC AGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAG GGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTAT GACGCTCTCCACATGCAAGCCCTGCCACC TAGGTAA AJ-ATGGCACTCCCCGTAACTGCTCTGCTGCT 143 MALPVTALLLPLALLL 144 21508CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEIVMTQSPATL HGCCCGGAAATAGTGATGACGCAGTCTCCA SVSPGERATLSCRASQGCCACCCTGTCTGTGTCTCCAGGGGAAAG SVSSNLAWYQQKPGQAAGCCACCCTCTCCTGCAGGGCCAGTCAGA PRLLIYGASTRATGIPGTGTTAGCAGCAACTTAGCCTGGTACCAG ARFSGSGSGTEFTLTICAGAAACCTGGCCAGGCTCCCAGGCTCCT SSLQSEDFAVYYCQQYCATCTATGGTGCATCCACCAGGGCCACTG AAYPTFGGGTKVEIKRGTATCCCAGCCAGGTTCAGTGGCAGTGGG GSTSGSGKPGSGEGSTTCTGGGACAGAGTTCACTCTCACCATCAG KGQVQLVQSGAEVKKPCAGCCTGCAGTCTGAAGATTTTGCAGTTT GASVKVSCKASGYTFTATTACTGTCAGCAGTACGCCGCCTACCCT SYYMHWVRQAPGQGLEACTTTTGGCGGAGGGACCAAGGTTGAGAT WMGIINPGGGSTSYAQCAAACGGGGGTCTACATCCGGCTCCGGGA KFQGRVTMTRDTSTSTAGCCCGGAAGTGGCGAAGGTAGTACAAAG VYMELSSLRSEDTAVYGGGCAGGTGCAGCTGGTGCAGTCTGGGGC YCARESWPMDVWGQGTTGAGGTGAAGAAGCCTGGGGCCTCAGTGA TVTVSSAAALDNEKSNAGGTTTCCTGCAAGGCATCTGGATACACC GTIIHVKGKHLCPSPLTTCACCAGCTACTATATGCACTGGGTGCG FPGPSKPFWVLVVVGGACAGGCCCCTGGACAAGGGCTTGAGTGGA VLACYSLLVTVAFIIFTGGGAATAATCAACCCTGGTGGTGGTAGC WVRSKRSRLLHSDYMNACAAGCTACGCACAGAAGTTCCAGGGCAG MTPRRPGPTRKHYQPYAGTCACCATGACCAGGGACACGTCCACGA APPRDFAAYRSRVKFSGCACAGTCTACATGGAGCTGAGCAGCCTG RSADAPAYQQGQNQLYAGATCTGAGGACACGGCGGTGTACTACTG NELNLGRREEYDVLDKCGCCAGAGAGAGTTGGCCAATGGACGTAT RRGRDPEMGGKPRRKNGGGGCCAGGGAACAACTGTCACCGTCTCC PQEGLYNELQKDKMAETCAGCCGCTGCCCTTGATAATGAAAAGTC AYSEIGMKGERRRGKGAAACGGAACAATCATTCACGTGAAGGGCA HDGLYQGLSTATKDTYAGCACCTCTGTCCGTCACCCTTGTTCCCT DALHMQALPPR GGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACT CTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCT GCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACAC TACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTT CCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCT CAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCT GAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGC AGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAG GGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTAT GACGCTCTCCACATGCAAGCCCTGCCACC TAGGTAA NM-ATGGCACTCCCCGTAACTGCTCTGCTGCT 145 MALPVTALLLPLALLL 146 21517CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQLQLQESGPGL LGCCCGCAGCTGCAGCTGCAGGAGTCGGGC VKPSETLSLTCTVSGGCCAGGACTGGTGAAGCCTTCGGAGACCCT SISSSSYYWGWIRQPPGTCCCTCACCTGCACTGTCTCTGGTGGCT GKGLEWIGSISYSGSTCCATCAGCAGTAGTAGTTACTACTGGGGC YYNPSLKSRVTISVDTTGGATCCGCCAGCCCCCAGGGAAGGGGCT SKNQFSLKLSSVTAADGGAGTGGATTGGGAGTATCTCCTATAGTG TAVYYCARGRGYATSLGGAGCACCTACTACAACCCGTCCCTCAAG AFDIWGQGTMVTVSSGAGTCGAGTCACCATATCCGTAGACACGTC STSGSGKPGSGEGSTKCAAGAACCAGTTCTCCCTGAAGCTGAGTT GEIVLTQSPATLSLSPCTGTGACCGCCGCAGACACGGCGGTGTAC GERATLSCRASQSVSSTACTGCGCCAGAGGCAGGGGATATGCAAC YLAWYQQKPGQAPRLLCAGCTTAGCCTTCGATATCTGGGGTCAGG IYDASNRATGIPARFSGTACAATGGTCACCGTCTCCTCAGGGTCT GSGSGTDFTLTISSLEACATCCGGCTCCGGGAAGCCCGGAAGTGG PEDFAVYYCQQRHVWPCGAAGGTAGTACAAAGGGGGAAATTGTGT PTFGGGTKVEIKRAAATGACACAGTCTCCAGCCACCCTGTCTTTG LDNEKSNGTIIHVKGKTCTCCAGGGGAAAGAGCCACCCTCTCCTG HLCPSPLFPGPSKPFWCAGGGCCAGTCAGAGTGTTAGCAGCTACT VLVVVGGVLACYSLLVTAGCCTGGTACCAACAGAAACCTGGCCAG TVAFIIFWVRSKRSRLGCTCCCAGGCTCCTCATCTATGATGCATC LHSDYMNMTPRRPGPTCAACAGGGCCACTGGCATCCCAGCCAGGT RKHYQPYAPPRDFAAYTCAGTGGCAGTGGGTCTGGGACAGACTTC RSRVKFSRSADAPAYQACTCTCACCATCAGCAGCCTAGAGCCTGA QGQNQLYNELNLGRREAGATTTTGCAGTTTATTACTGTCAGCAGA EYDVLDKRRGRDPEMGGACACGTCTGGCCTCCTACTTTTGGCGGA GKPRRKNPQEGLYNELGGGACCAAGGTTGAGATCAAACGGGCCGC QKDKMAEAYSEIGMKGTGCCCTTGATAATGAAAAGTCAAACGGAA ERRRGKGHDGLYQGLSCAATCATTCACGTGAAGGGCAAGCACCTC TATKDTYDALHMQALPTGTCCGTCACCCTTGTTCCCTGGTCCATC PR CAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTC GTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATA GCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCC TTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCT GCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGG GACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGG TGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGAT AAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAG GGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCT CCACATGCAAGCCCTGCCACCTAGGTAA NM-ATGGCACTCCCCGTAACTGCTCTGCTGCT 147 MALPVTALLLPLALLL 148 21517CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEIVLTQSPATL HGCCCGGAAATTGTGTTGACACAGTCTCCA SLSPGERATLSCRASQGCCACCCTGTCTTTGTCTCCAGGGGAAAG SVSSYLAWYQQKPGQAAGCCACCCTCTCCTGCAGGGCCAGTCAGA PRLLIYDASNRATGIPGTGTTAGCAGCTACTTAGCCTGGTACCAA ARFSGSGSGTDFTLTICAGAAACCTGGCCAGGCTCCCAGGCTCCT SSLEPEDFAVYYCQQRCATCTATGATGCATCCAACAGGGCCACTG HVWPPTFGGGTKVEIKGCATCCCAGCCAGGTTCAGTGGCAGTGGG RGSTSGSGKPGSGEGSTCTGGGACAGACTTCACTCTCACCATCAG TKGQLQLQESGPGLVKCAGCCTAGAGCCTGAAGATTTTGCAGTTT PSETLSLTCTVSGGSIATTACTGTCAGCAGAGACACGTCTGGCCT SSSSYYWGWIRQPPGKCCTACTTTTGGCGGAGGGACCAAGGTTGA GLEWIGSISYSGSTYYGATCAAACGGGGGTCTACATCCGGCTCCG NPSLKSRVTISVDTSKGGAAGCCCGGAAGTGGCGAAGGTAGTACA NQFSLKLSSVTAADTAAAGGGGCAGCTGCAGCTGCAGGAGTCGGG VYYCARGRGYATSLAFCCCAGGACTGGTGAAGCCTTCGGAGACCC DIWGQGTMVTVSSAAATGTCCCTCACCTGCACTGTCTCTGGTGGC LDNEKSNGTIIHVKGKTCCATCAGCAGTAGTAGTTACTACTGGGG HLCPSPLFPGPSKPFWCTGGATCCGCCAGCCCCCAGGGAAGGGGC VLVVVGGVLACYSLLVTGGAGTGGATTGGGAGTATCTCCTATAGT TVAFIIFWVRSKRSRLGGGAGCACCTACTACAACCCGTCCCTCAA LHSDYMNMTPRRPGPTGAGTCGAGTCACCATATCCGTAGACACGT RKHYQPYAPPRDFAAYCCAAGAACCAGTTCTCCCTGAAGCTGAGT RSRVKFSRSADAPAYQTCTGTGACCGCCGCAGACACGGCGGTGTA QGQNQLYNELNLGRRECTACTGCGCCAGAGGCAGGGGATATGCAA EYDVLDKRRGRDPEMGCCAGCTTAGCCTTCGATATCTGGGGTCAG GKPRRKNPQEGLYNELGGTACAATGGTCACCGTCTCCTCAGCCGC QKDKMAEAYSEIGMKGTGCCCTTGATAATGAAAAGTCAAACGGAA ERRRGKGHDGLYQGLSCAATCATTCACGTGAAGGGCAAGCACCTC TATKDTYDALHMQALPTGTCCGTCACCCTTGTTCCCTGGTCCATC PR CAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTC GTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATA GCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCC TTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCT GCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGG GACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGG TGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGAT AAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAG GGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCT CCACATGCAAGCCCTGCCACCTAGGTAA TS-ATGGCACTCCCCGTAACTGCTCTGCTGCT 149 MALPVTALLLPLALLL 150 21522CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEVQLVESGGGL LGCCCGGAGGTGCAGCTGGTGGAGTCTGGG VQPGGSLRLSCAASGFGGAGGCTTGGTACAGCCTGGGGGGTCCCT TFSSYSMNWVRQAPGKGAGACTCTCCTGTGCAGCCTCTGGATTCA GLEWVSTISSSSSTIYCCTTCAGTAGCTATAGCATGAACTGGGTC YADSVKGRFTISRDNACGCCAGGCTCCAGGGAAGGGGCTGGAGTG KNSLYLQMNSLRAEDTGGTTTCAACCATTAGTAGTAGTAGTAGTA AVYYCARGSQEHLIFDCCATATACTACGCAGACTCTGTGAAGGGC YWGQGTLVTVSSGSTSCGATTCACCATCTCCAGAGACAATGCCAA GSGKPGSGEGSTKGEIGAACTCACTGTATCTGCAAATGAACAGCC VLTQSPATLSLSPGERTGAGAGCTGAGGACACGGCGGTGTACTAC ATLSCRASQSVSRYLATGCGCCAGAGGTTCTCAGGAGCACCTGAT WYQQKPGQAPRLLIYDTTTCGATTATTGGGGACAGGGTACATTGG ASNRATGIPARFSGSGTCACCGTCTCCTCAGGGTCTACATCCGGC SGTDFTLTISSLEPEDTCCGGGAAGCCCGGAAGTGGCGAAGGTAG FAVYYCQQRFYYPWTFTACAAAGGGGGAAATTGTGTTGACACAGT GGGTKVEIKRAAALDNCTCCAGCCACCCTGTCTTTGTCTCCAGGG EKSNGTIIHVKGKHLCGAAAGAGCCACCCTCTCCTGCAGGGCCAG PSPLFPGPSKPFWVLVTCAGAGTGTTAGCAGGTACTTAGCCTGGT VVGGVLACYSLLVTVAACCAACAGAAACCTGGCCAGGCTCCCAGG FIIFWVRSKRSRLLHSCTCCTCATCTATGATGCATCCAACAGGGC DYMNMTPRRPGPTRKHCACTGGCATCCCAGCCAGGTTCAGTGGCA YQPYAPPRDFAAYRSRGTGGGTCTGGGACAGACTTCACTCTCACC VKFSRSADAPAYQQGQATCAGCAGCCTAGAGCCTGAAGATTTTGC NQLYNELNLGRREEYDAGTTTATTACTGTCAGCAGAGATTCTACT VLDKRRGRDPEMGGKPACCCTTGGACTTTTGGCGGAGGGACCAAG RRKNPQEGLYNELQKDGTTGAGATCAAACGGGCCGCTGCCCTTGA KMAEAYSEIGMKGERRTAATGAAAAGTCAAACGGAACAATCATTC RGKGHDGLYQGLSTATACGTGAAGGGCAAGCACCTCTGTCCGTCA KDTYDALHMQALPPRCCCTTGTTCCCTGGTCCATCCAAGCCATT CTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTG GCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACA TGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACC ACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCA CCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGG AAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACC AAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCT GAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACG GTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCA AGCCCTGCCACCTAGGTAA TS-ATGGCACTCCCCGTAACTGCTCTGCTGCT 151 MALPVTALLLPLALLL 152 21522CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEIVLTQSPATL HGCCCGGAAATTGTGTTGACACAGTCTCCA SLSPGERATLSCRASQGCCACCCTGTCTTTGTCTCCAGGGGAAAG SVSRYLAWYQQKPGQAAGCCACCCTCTCCTGCAGGGCCAGTCAGA PRLLIYDASNRATGIPGTGTTAGCAGGTACTTAGCCTGGTACCAA ARFSGSGSGTDFTLTICAGAAACCTGGCCAGGCTCCCAGGCTCCT SSLEPEDFAVYYCQQRCATCTATGATGCATCCAACAGGGCCACTG FYYPWTFGGGTKVEIKGCATCCCAGCCAGGTTCAGTGGCAGTGGG RGSTSGSGKPGSGEGSTCTGGGACAGACTTCACTCTCACCATCAG TKGEVQLVESGGGLVQCAGCCTAGAGCCTGAAGATTTTGCAGTTT PGGSLRLSCAASGFTFATTACTGTCAGCAGAGATTCTACTACCCT SSYSMNWVRQAPGKGLTGGACTTTTGGCGGAGGGACCAAGGTTGA EWVSTISSSSSTIYYAGATCAAACGGGGGTCTACATCCGGCTCCG DSVKGRFTISRDNAKNGGAAGCCCGGAAGTGGCGAAGGTAGTACA SLYLQMNSLRAEDTAVAAGGGGGAGGTGCAGCTGGTGGAGTCTGG YYCARGSQEHLIFDYWGGGAGGCTTGGTACAGCCTGGGGGGTCCC GQGTLVTVSSAAALDNTGAGACTCTCCTGTGCAGCCTCTGGATTC EKSNGTIIHVKGKHLCACCTTCAGTAGCTATAGCATGAACTGGGT PSPLFPGPSKPFWVLVCCGCCAGGCTCCAGGGAAGGGGCTGGAGT VVGGVLACYSLLVTVAGGGTTTCAACCATTAGTAGTAGTAGTAGT FIIFWVRSKRSRLLHSACCATATACTACGCAGACTCTGTGAAGGG DYMNMTPRRPGPTRKHCCGATTCACCATCTCCAGAGACAATGCCA YQPYAPPRDFAAYRSRAGAACTCACTGTATCTGCAAATGAACAGC VKFSRSADAPAYQQGQCTGAGAGCTGAGGACACGGCGGTGTACTA NQLYNELNLGRREEYDCTGCGCCAGAGGTTCTCAGGAGCACCTGA VLDKRRGRDPEMGGKPTTTTCGATTATTGGGGACAGGGTACATTG RRKNPQEGLYNELQKDGTCACCGTCTCCTCAGCCGCTGCCCTTGA KMAEAYSEIGMKGERRTAATGAAAAGTCAAACGGAACAATCATTC RGKGHDGLYQGLSTATACGTGAAGGGCAAGCACCTCTGTCCGTCA KDTYDALHMQALPPRCCCTTGTTCCCTGGTCCATCCAAGCCATT CTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTG GCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACA TGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACC ACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCA CCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGG AAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACC AAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCT GAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACG GTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCA AGCCCTGCCACCTAGGTAA RY-ATGGCACTCCCCGTAACTGCTCTGCTGCT 153 MALPVTALLLPLALLL 154 21527CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVESGGGV LGCCCGCAGGTGCAGCTGGTGGAGTCTGGG VQPGRSLRLSCAASGFGGAGGCGTGGTCCAGCCTGGGAGGTCCCT TFSSYGMHWVRQAPGKGAGACTCTCCTGTGCAGCGTCTGGATTCA GLEWVAVISYDGSNKYCCTTCAGTAGCTATGGCATGCACTGGGTC YADSVKGRFTISRDNSCGCCAGGCTCCAGGCAAGGGGCTGGAGTG KNTLYLQMNSLRAEDTGGTGGCAGTTATATCGTATGATGGAAGTA AVYYCARTDFWSGSPPATAAATACTATGCAGACTCCGTGAAGGGC GLDYWGQGTLVTVSSGCGATTCACCATCTCCAGAGACAATTCCAA STSGSGKPGSGEGSTKGAACACGCTGTATCTGCAAATGAACAGCC GDIQLTQSPSSVSASVTGAGAGCCGAGGACACGGCGGTGTACTAC GDRVTITCRASQGISSTGCGCCAGAACTGACTTCTGGAGCGGATC WLAWYQQKPGKAPKLLCCCTCCAGGCTTAGATTACTGGGGACAGG IYGASSLQSGVPSRFSGTACATTGGTCACCGTCTCCTCAGGGTCT GSGSGTDFTLTISSLQACATCCGGCTCCGGGAAGCCCGGAAGTGG PEDFATYYCQQIYTFPCGAAGGTAGTACAAAGGGGGACATCCAGT FTFGGGTKVEIKRAAATGACCCAGTCTCCATCTTCCGTGTCTGCA LDNEKSNGTIIHVKGKTCTGTAGGAGACAGAGTCACCATCACTTG HLCPSPLFPGPSKPFWTCGGGCGAGTCAGGGTATTAGCAGCTGGT VLVVVGGVLACYSLLVTAGCCTGGTATCAGCAGAAACCAGGGAAA TVAFIIFWVRSKRSRLGCCCCTAAGCTCCTGATCTATGGTGCATC LHSDYMNMTPRRPGPTCAGTTTGCAAAGTGGGGTCCCATCAAGGT RKHYQPYAPPRDFAAYTCAGCGGCAGTGGATCTGGGACAGATTTC RSRVKFSRSADAPAYQACTCTCACCATCAGCAGCCTGCAGCCTGA QGQNQLYNELNLGRREAGATTTTGCAACTTATTACTGTCAGCAGA EYDVLDKRRGRDPEMGTATACACCTTCCCTTTCACTTTTGGCGGA GKPRRKNPQEGLYNELGGGACCAAGGTTGAGATCAAACGGGCCGC QKDKMAEAYSEIGMKGTGCCCTTGATAATGAAAAGTCAAACGGAA ERRRGKGHDGLYQGLSCAATCATTCACGTGAAGGGCAAGCACCTC TATKDTYDALHMQALPTGTCCGTCACCCTTGTTCCCTGGTCCATC PR CAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTC GTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATA GCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCC TTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCT GCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGG GACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGG TGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGAT AAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAG GGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCT CCACATGCAAGCCCTGCCACCTAGGTAA RY-ATGGCACTCCCCGTAACTGCTCTGCTGCT 155 MALPVTALLLPLALLL 156 21527CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPDIQLTQSPSSV HGCCCGGACATCCAGTTGACCCAGTCTCCA SASVGDRVTITCRASQTCTTCCGTGTCTGCATCTGTAGGAGACAG GISSWLAWYQQKPGKAAGTCACCATCACTTGTCGGGCGAGTCAGG PKLLIYGASSLQSGVPGTATTAGCAGCTGGTTAGCCTGGTATCAG SRFSGSGSGTDFTLTICAGAAACCAGGGAAAGCCCCTAAGCTCCT SSLQPEDFATYYCQQIGATCTATGGTGCATCCAGTTTGCAAAGTG YTFPFTFGGGTKVEIKGGGTCCCATCAAGGTTCAGCGGCAGTGGA RGSTSGSGKPGSGEGSTCTGGGACAGATTTCACTCTCACCATCAG TKGQVQLVESGGGVVQCAGCCTGCAGCCTGAAGATTTTGCAACTT PGRSLRLSCAASGFTFATTACTGTCAGCAGATATACACCTTCCCT SSYGMHWVRQAPGKGLTTCACTTTTGGCGGAGGGACCAAGGTTGA EWVAVISYDGSNKYYAGATCAAACGGGGGTCTACATCCGGCTCCG DSVKGRFTISRDNSKNGGAAGCCCGGAAGTGGCGAAGGTAGTACA TLYLQMNSLRAEDTAVAAGGGGCAGGTGCAGCTGGTGGAGTCTGG YYCARTDFWSGSPPGLGGGAGGCGTGGTCCAGCCTGGGAGGTCCC DYWGQGTLVTVSSAAATGAGACTCTCCTGTGCAGCGTCTGGATTC LDNEKSNGTIIHVKGKACCTTCAGTAGCTATGGCATGCACTGGGT HLCPSPLFPGPSKPFWCCGCCAGGCTCCAGGCAAGGGGCTGGAGT VLVVVGGVLACYSLLVGGGTGGCAGTTATATCGTATGATGGAAGT TVAFIIFWVRSKRSRLAATAAATACTATGCAGACTCCGTGAAGGG LHSDYMNMTPRRPGPTCCGATTCACCATCTCCAGAGACAATTCCA RKHYQPYAPPRDFAAYAGAACACGCTGTATCTGCAAATGAACAGC RSRVKFSRSADAPAYQCTGAGAGCCGAGGACACGGCGGTGTACTA QGQNQLYNELNLGRRECTGCGCCAGAACTGACTTCTGGAGCGGAT EYDVLDKRRGRDPEMGCCCCTCCAGGCTTAGATTACTGGGGACAG GKPRRKNPQEGLYNELGGTACATTGGTCACCGTCTCCTCAGCCGC QKDKMAEAYSEIGMKGTGCCCTTGATAATGAAAAGTCAAACGGAA ERRRGKGHDGLYQGLSCAATCATTCACGTGAAGGGCAAGCACCTC TATKDTYDALHMQALPTGTCCGTCACCCTTGTTCCCTGGTCCATC PR CAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTC GTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATA GCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCC TTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCT GCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGG GACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGG TGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGAT AAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAG GGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCT CCACATGCAAGCCCTGCCACCTAGGTAA PP-ATGGCACTCCCCGTAACTGCTCTGCTGCT 157 MALPVTALLLPLALLL 158 21528CARHxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVQSGAEV LGCCCGCAGGTGCAGCTGGTGCAGTCTGGG KKPGSSVKVSCKASGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGT TFSSYAISWVRQAPGQGAAGGTCTCCTGCAAGGCTTCTGGAGGCA GLEWMGGIIPIFGTANCCTTCAGCAGCTATGCTATCAGCTGGGTG YAQKFQGRVTITADESCGACAGGCCCCTGGACAAGGGCTTGAGTG TSTAYMELSSLRSEDTGATGGGAGGGATCATCCCTATCTTTGGTA AVYYCARTPEYSSSIWCAGCAAACTACGCACAGAAGTTCCAGGGC HYYYGMDVWGQGTTVTAGAGTCACGATTACCGCGGACGAATCCAC VSSGSTSGSGKPGSGEGAGCACAGCCTACATGGAGCTGAGCAGCC GSTKGDIVMTQSPDSLTGAGATCTGAGGACACGGCGGTGTACTAC AVSLGERATINCKSSQTGCGCCAGAACTCCTGAATACTCCTCCAG SVLYSSNNKNYLAWYQCATATGGCACTATTACTACGGCATGGACG QKPGQPPKLLIYWASTTATGGGGCCAGGGAACAACTGTCACCGTC RESGVPDRFSGSGSGTTCCTCAGGGTCTACATCCGGCTCCGGGAA DFTLTISSLQAEDVAVGCCCGGAAGTGGCGAAGGTAGTACAAAGG YYCQQFAHTPFTFGGGGGGACATCGTGATGACCCAGTCTCCAGAC TKVEIKRAAALDNEKSTCCCTGGCTGTGTCTCTGGGCGAGAGGGC NGTIIHVKGKHLCPSPCACCATCAACTGCAAGTCCAGCCAGAGTG LFPGPSKPFWVLVVVGTTTTATACAGCTCCAACAATAAGAACTAC GVLACYSLLVTVAFIITTAGCTTGGTACCAGCAGAAACCAGGACA FWVRSKRSRLLHSDYMGCCTCCTAAGCTGCTCATTTACTGGGCAT NMTPRRPGPTRKHYQPCTACCCGGGAATCCGGGGTCCCTGACCGA YAPPRDFAAYRSRVKFTTCAGTGGCAGCGGGTCTGGGACAGATTT SRSADAPAYQQGQNQLCACTCTCACCATCAGCAGCCTGCAGGCTG YNELNLGRREEYDVLDAAGATGTGGCAGTTTATTACTGTCAGCAG KRRGRDPEMGGKPRRKTTCGCCCACACTCCTTTCACTTTTGGCGG NPQEGLYNELQKDKMAAGGGACCAAGGTTGAGATCAAACGGGCCG EAYSEIGMKGERRRGKCTGCCCTTGATAATGAAAAGTCAAACGGA GHDGLYQGLSTATKDTACAATCATTCACGTGAAGGGCAAGCACCT YDALHMQALPPR CTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTG GGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGG TTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCG CCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCC TATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCC AGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGA CAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAG GAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAG GCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAG CACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA PP- ATGGCACTCCCCGTAACTGCTCTGCTGCT 159MALPVTALLLPLALLL 160 21528CARLx GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPDIVMTQSPDSL H GCCCGGACATCGTGATGACCCAGTCTCCA AVSLGERATINCKSSQGACTCCCTGGCTGTGTCTCTGGGCGAGAG SVLYSSNNKNYLAWYQGGCCACCATCAACTGCAAGTCCAGCCAGA QKPGQPPKLLIYWASTGTGTTTTATACAGCTCCAACAATAAGAAC RESGVPDRFSGSGSGTTACTTAGCTTGGTACCAGCAGAAACCAGG DFTLTISSLQAEDVAVACAGCCTCCTAAGCTGCTCATTTACTGGG YYCQQFAHTPFTFGGGCATCTACCCGGGAATCCGGGGTCCCTGAC TKVEIKRGSTSGSGKPCGATTCAGTGGCAGCGGGTCTGGGACAGA GSGEGSTKGQVQLVQSTTTCACTCTCACCATCAGCAGCCTGCAGG GAEVKKPGSSVKVSCKCTGAAGATGTGGCAGTTTATTACTGTCAG ASGGTFSSYAISWVRQCAGTTCGCCCACACTCCTTTCACTTTTGG APGQGLEWMGGIIPIFCGGAGGGACCAAGGTTGAGATCAAACGGG GTANYAQKFQGRVTITGGTCTACATCCGGCTCCGGGAAGCCCGGA ADESTSTAYMELSSLRAGTGGCGAAGGTAGTACAAAGGGGCAGGT SEDTAVYYCARTPEYSGCAGCTGGTGCAGTCTGGGGCTGAGGTGA SSIWHYYYGMDVWGQGAGAAGCCTGGGTCCTCGGTGAAGGTCTCC TTVTVSSAAALDNEKSTGCAAGGCTTCTGGAGGCACCTTCAGCAG NGTIIHVKGKHLCPSPCTATGCTATCAGCTGGGTGCGACAGGCCC LFPGPSKPFWVLVVVGCTGGACAAGGGCTTGAGTGGATGGGAGGG GVLACYSLLVTVAFIIATCATCCCTATCTTTGGTACAGCAAACTA FWVRSKRSRLLHSDYMCGCACAGAAGTTCCAGGGCAGAGTCACGA NMTPRRPGPTRKHYQPTTACCGCGGACGAATCCACGAGCACAGCC YAPPRDFAAYRSRVKFTACATGGAGCTGAGCAGCCTGAGATCTGA SRSADAPAYQQGQNQLGGACACGGCGGTGTACTACTGCGCCAGAA YNELNLGRREEYDVLDCTCCTGAATACTCCTCCAGCATATGGCAC KRRGRDPEMGGKPRRKTATTACTACGGCATGGACGTATGGGGCCA NPQEGLYNELQKDKMAGGGAACAACTGTCACCGTCTCCTCAGCCG EAYSEIGMKGERRRGKCTGCCCTTGATAATGAAAAGTCAAACGGA GHDGLYQGLSTATKDTACAATCATTCACGTGAAGGGCAAGCACCT YDALHMQALPPR CTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTG GGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGG TTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCG CCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCC TATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCC AGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGA CAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAG GAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAG GCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAG CACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA RD- ATGGCACTCCCCGTAACTGCTCTGCTGCT 161MALPVTALLLPLALLL 162 21530CARHx GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPQVQLVESGGGV L GCCCGCAGGTGCAGCTGGTGGAGTCTGGG VQPGRSLRLSCAASGFGGAGGCGTGGTCCAGCCTGGGAGGTCCCT TFSSYGMHWVRQAPGKGAGACTCTCCTGTGCAGCGTCTGGATTCA GLEWVAVISYDGSNKYCCTTCAGTAGCTATGGCATGCACTGGGTC YADSVKGRFTISRDNSCGCCAGGCTCCAGGCAAGGGGCTGGAGTG KNTLYLQMNSLRAEDTGGTGGCAGTTATATCGTATGATGGAAGTA AVYYCVKGPLQEPPYDATAAATACTATGCAGACTCCGTGAAGGGC YGMDVWGQGTTVTVSSCGATTCACCATCTCCAGAGACAATTCCAA GSTSGSGKPGSGEGSTGAACACGCTGTATCTGCAAATGAACAGCC KGEIVMTQSPATLSVSTGAGAGCCGAGGACACGGCGGTGTACTAC PGERATLSCRASQSVSTGCGTCAAGGGGCCGTTGCAGGAGCCGCC SNLAWYQQKPGQAPRLATACGATTATGGAATGGACGTATGGGGCC LIYSASTRATGIPARFAGGGAACAACTGTCACCGTCTCCTCAGGG SGSGSGTEFTLTISSLTCTACATCCGGCTCCGGGAAGCCCGGAAG QSEDFAVYYCQQHHVWTGGCGAAGGTAGTACAAAGGGGGAAATAG PLTFGGGTKVEIKRAATGATGACGCAGTCTCCAGCCACCCTGTCT ALDNEKSNGTIIHVKGGTGTCTCCAGGGGAAAGAGCCACCCTCTC KHLCPSPLFPGPSKPFCTGCAGGGCCAGTCAGAGTGTTAGCAGCA WVLVVVGGVLACYSLLACTTAGCCTGGTACCAGCAGAAACCTGGC VTVAFIIFWVRSKRSRCAGGCTCCCAGGCTCCTCATCTATAGCGC LLHSDYMNMTPRRPGPATCCACCAGGGCCACTGGTATCCCAGCCA TRKHYQPYAPPRDFAAGGTTCAGTGGCAGTGGGTCTGGGACAGAG YRSRVKFSRSADAPAYTTCACTCTCACCATCAGCAGCCTGCAGTC QQGQNQLYNELNLGRRTGAAGATTTTGCAGTTTATTACTGTCAGC EEYDVLDKRRGRDPEMAGCACCACGTCTGGCCTCTCACTTTTGGC GGKPRRKNPQEGLYNEGGAGGGACCAAGGTTGAGATCAAACGGGC LQKDKMAEAYSEIGMKCGCTGCCCTTGATAATGAAAAGTCAAACG GERRRGKGHDGLYQGLGAACAATCATTCACGTGAAGGGCAAGCAC STATKDTYDALHMQALCTCTGTCCGTCACCCTTGTTCCCTGGTCC PPR ATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTG CTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCC ATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCA GCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGA TCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACC TGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGAT GGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAG GATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAA AAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGC TCTCCACATGCAAGCCCTGCCACCTAGGT AA RD-ATGGCACTCCCCGTAACTGCTCTGCTGCT 163 MALPVTALLLPLALLL 164 21530CARLxGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPEIVMTQSPATL HGCCCGGAAATAGTGATGACGCAGTCTCCA SVSPGERATLSCRASQGCCACCCTGTCTGTGTCTCCAGGGGAAAG SVSSNLAWYQQKPGQAAGCCACCCTCTCCTGCAGGGCCAGTCAGA PRLLIYSASTRATGIPGTGTTAGCAGCAACTTAGCCTGGTACCAG ARFSGSGSGTEFTLTICAGAAACCTGGCCAGGCTCCCAGGCTCCT SSLQSEDFAVYYCQQHCATCTATAGCGCATCCACCAGGGCCACTG HVWPLTFGGGTKVEIKGTATCCCAGCCAGGTTCAGTGGCAGTGGG RGSTSGSGKPGSGEGSTCTGGGACAGAGTTCACTCTCACCATCAG TKGQVQLVESGGGVVQCAGCCTGCAGTCTGAAGATTTTGCAGTTT PGRSLRLSCAASGFTFATTACTGTCAGCAGCACCACGTCTGGCCT SSYGMHWVRQAPGKGLCTCACTTTTGGCGGAGGGACCAAGGTTGA EWVAVISYDGSNKYYAGATCAAACGGGGGTCTACATCCGGCTCCG DSVKGRFTISRDNSKNGGAAGCCCGGAAGTGGCGAAGGTAGTACA TLYLQMNSLRAEDTAVAAGGGGCAGGTGCAGCTGGTGGAGTCTGG YYCVKGPLQEPPYDYGGGGAGGCGTGGTCCAGCCTGGGAGGTCCC MDVWGQGTTVTVSSAATGAGACTCTCCTGTGCAGCGTCTGGATTC ALDNEKSNGTIIHVKGACCTTCAGTAGCTATGGCATGCACTGGGT KHLCPSPLFPGPSKPFCCGCCAGGCTCCAGGCAAGGGGCTGGAGT WVLVVVGGVLACYSLLGGGTGGCAGTTATATCGTATGATGGAAGT VTVAFIIFWVRSKRSRAATAAATACTATGCAGACTCCGTGAAGGG LLHSDYMNMTPRRPGPCCGATTCACCATCTCCAGAGACAATTCCA TRKHYQPYAPPRDFAAAGAACACGCTGTATCTGCAAATGAACAGC YRSRVKFSRSADAPAYCTGAGAGCCGAGGACACGGCGGTGTACTA QQGQNQLYNELNLGRRCTGCGTCAAGGGGCCGTTGCAGGAGCCGC EEYDVLDKRRGRDPEMCATACGATTATGGAATGGACGTATGGGGC GGKPRRKNPQEGLYNECAGGGAACAACTGTCACCGTCTCCTCAGC LQKDKMAEAYSEIGMKCGCTGCCCTTGATAATGAAAAGTCAAACG GERRRGKGHDGLYQGLGAACAATCATTCACGTGAAGGGCAAGCAC STATKDTYDALHMQALCTCTGTCCGTCACCCTTGTTCCCTGGTCC PPR ATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTG CTCGTCACCGTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCC ATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCA GCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGA TCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACC TGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGAT GGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAG GATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAA AAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGC TCTCCACATGCAAGCCCTGCCACCTAGGT AAClone 24C1 ATGGCACTCCCCGTAACTGCTCTGCTGCT 165 MALPVTALLLPLALLL 166THD CAR GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL DNA HxLGCCCGCAGGTCCAACTGCAAGAAAGCGGA VKPSETLSLTCTVSGGCCCGGACTGGTGAAGCCTTCTGAGACACT SISSYYWSWIRQPPGKTAGTCTGACGTGCACGGTCAGTGGCGGCT GLEWIGYIYYSGSTNYCCATCTCCTCCTATTATTGGTCATGGATA NPSLKSRVTISVDTSKCGACAACCCCCAGGTAAGGGCCTGGAATG NQFSLKLSSVTAADTAGATTGGCTATATCTACTATTCAGGAAGCA VYYCVSLVYCGGDCYSCGAACTACAATCCCAGCCTGAAGTCCCGA GFDYWGQGTLVTVSSGGTGACAATTTCAGTAGATACCAGTAAAAA GGGSGGGGSGGGGSDICCAGTTCAGTCTTAAACTGTCAAGCGTGA QLTQSPSSLSASVGDRCAGCTGCCGACACCGCTGTGTATTACTGC VSFTCQASQDINNFLNGTCTCACTGGTGTATTGTGGAGGGGATTG WYQQKPGKAPKLLIYDTTATAGCGGGTTCGATTATTGGGGACAGG ASNLETGVPSRFSGSGGAACCCTGGTGACTGTATCTTCCGGCGGC SGTDFTFTISSLQPEDGGCGGCTCAGGGGGTGGCGGTAGTGGCGG IATYYCQQYGNLPFTFTGGGGGTTCCGATATTCAACTGACACAAT GGGTKVEIKRAAALDNCCCCCAGCTCACTCAGCGCCAGCGTGGGG EKSNGTIIHVKGKHLCGACAGGGTTAGCTTTACCTGTCAAGCCTC PSPLFPGPSKPFWVLVTCAGGATATAAATAACTTTCTGAACTGGT VVGGVLACYSLLVTVAATCAACAGAAGCCTGGGAAGGCGCCCAAA FIIFWVRSKRSRLLHSCTCCTGATCTATGATGCGTCCAACCTGGA DYMNMTPRRPGPTRKHAACTGGCGTGCCTTCACGCTTTAGCGGCT YQPYAPPRDFAAYRSRCTGGCAGTGGTACAGACTTCACTTTTACC VKFSRSADAPAYQQGQATCTCTTCACTTCAGCCGGAGGACATCGC NQLYNELNLGRREEYDCACATATTACTGTCAACAGTACGGAAACT VLDKRRGRDPEMGGKPTGCCCTTTACTTTTGGAGGCGGCACCAAA RRKNPQEGLYNELQKDGTTGAAATCAAAAGGGCCGCTGCCCTGGA KMAEAYSEIGMKGERRTAACGAAAAGAGCAATGGGACTATAATAC RGKGHDGLYQGLSTATATGTTAAAGGAAAACACCTGTGTCCATCT KDTYDALHMQALPPRCCCCTGTTCCCTGGACCGTCAAAGCCATT TTGGGTGCTCGTGGTTGTCGGTGGCGTTCTCGCCTGTTATAGCTTGCTGGTGACAGTA GCCTTCATTATCTTTTGGGTGAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACA TGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACC ACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCA CCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGG AAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACC AAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCT GAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACG GTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCA AGCCCTGCCACCTAGGTAA (CAR1.1)CAGGTCCAACTGCAAGAAAGCGGACCCGG 167 QVQLQESGPGLVKPSE 168 Clone 24C1ACTGGTGAAGCCTTCTGAGACACTTAGTC TLSLTCTVSGGSISSY THD CARTGACGTGCACGGTCAGTGGCGGCTCCATC YWSWIRQPPGKGLEWI DNA HxLTCCTCCTATTATTGGTCATGGATACGACA GYIYYSGSTNYNPSLKACCCCCAGGTAAGGGCCTGGAATGGATTG SRVTISVDTSKNQFSLGCTATATCTACTATTCAGGAAGCACGAAC KLSSVTAADTAVYYCVTACAATCCCAGCCTGAAGTCCCGAGTGAC SLVYCGGDCYSGFDYWAATTTCAGTAGATACCAGTAAAAACCAGT GQGTLVTVSSGGGGSGTCAGTCTTAAACTGTCAAGCGTGACAGCT GGGSGGGGSDIQLTQSGCCGACACCGCTGTGTATTACTGCGTCTC PSSLSASVGDRVSFTCACTGGTGTATTGTGGAGGGGATTGTTATA QASQDINNFLNWYQQKGCGGGTTCGATTATTGGGGACAGGGAACC PGKAPKLLIYDASNLECTGGTGACTGTATCTTCCGGCGGCGGCGG TGVPSRFSGSGSGTDFCTCAGGGGGTGGCGGTAGTGGCGGTGGGG TFTISSLQPEDIATYYGTTCCGATATTCAACTGACACAATCCCCC CQQYGNLPFTFGGGTKAGCTCACTCAGCGCCAGCGTGGGGGACAG VEIKRAAALDNEKSNGGGTTAGCTTTACCTGTCAAGCCTCTCAGG TIIHVKGKHLCPSPLFATATAAATAACTTTCTGAACTGGTATCAA PGPSKPFWVLVVVGGVCAGAAGCCTGGGAAGGCGCCCAAACTCCT LACYSLLVTVAFIIFWGATCTATGATGCGTCCAACCTGGAAACTG VRSKRSRLLHSDYMNMGCGTGCCTTCACGCTTTAGCGGCTCTGGC TPRRPGPTRKHYQPYAAGTGGTACAGACTTCACTTTTACCATCTC PPRDFAAYRSRVKFSRTTCACTTCAGCCGGAGGACATCGCCACAT SADAPAYQQGQNQLYNATTACTGTCAACAGTACGGAAACTTGCCC ELNLGRREEYDVLDKRTTTACTTTTGGAGGCGGCACCAAAGTTGA RGRDPEMGGKPRRKNPAATCAAAAGGGCCGCTGCCCTGGATAACG QEGLYNELQKDKMAEAAAAAGAGCAATGGGACTATAATACATGTT YSEIGMKGERRRGKGHAAAGGAAAACACCTGTGTCCATCTCCCCT DGLYQGLSTATKDTYDGTTCCCTGGACCGTCAAAGCCATTTTGGG ALHMQALPPR TGCTCGTGGTTGTCGGTGGCGTTCTCGCCTGTTATAGCTTGCTGGTGACAGTAGCCTT CATTATCTTTTGGGTGAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAAT ATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTA GAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGC GTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAG TATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGAC GAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGC CTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTG TACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCC TGCCACCTAGG (CAR1.2)ATGGCACTCCCCGTAACTGCTCTGCTGCT 169 MALPVTALLLPLALLL 170 Clone 24C1GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL CHD CARGCCCGCAGGTGCAGCTGCAGGAATCCGGA VKPSETLSLTCTVSGG DNA HxLCCGGGGCTGGTGAAGCCCAGCGAGACTCT SISSYYWSWIRQPPGKGAGTCTCACGTGTACAGTTTCTGGAGGTA GLEWIGYIYYSGSTNYGCATTAGCTCCTACTATTGGTCATGGATA NPSLKSRVTISVDTSKAGGCAGCCCCCCGGGAAGGGATTGGAATG NQFSLKLSSVTAADTAGATCGGCTATATTTACTACAGTGGGAGCA VYYCVSLVYCGGDCYSCCAATTACAACCCCTCACTGAAGTCTAGA GFDYWGQGTLVTVSSGGTTACAATCAGCGTTGACACCTCAAAGAA GGGSGGGGSGGGGSDITCAGTTCAGTTTGAAATTGTCTAGCGTCA QLTQSPSSLSASVGDRCAGCAGCTGATACAGCCGTCTATTATTGT VSFTCQASQDINNFLNGTTTCTCTGGTCTATTGCGGTGGGGATTG WYQQKPGKAPKLLIYDTTACAGTGGCTTTGACTATTGGGGGCAGG ASNLETGVPSRFSGSGGTACTCTGGTTACAGTTTCTTCCGGGGGG SGTDFTFTISSLQPEDGGAGGCTCTGGGGGCGGAGGCTCAGGTGG IATYYCQQYGNLPFTFTGGAGGCAGCGACATCCAGTTGACACAGA GGGTKVEIKRAAAIEVGCCCGAGTTCCTTGTCCGCCTCCGTCGGG MYPPPYLDNEKSNGTIGATAGAGTGTCATTTACCTGTCAGGCCTC IHVKGKHLCPSPLFPGTCAGGATATTAATAACTTTCTGAATTGGT PSKPFWVLVVVGGVLAATCAGCAAAAGCCCGGAAAGGCACCCAAG CYSLLVTVAFIIFWVRCTGTTGATTTACGACGCCAGTAACCTGGA SKRSRLLHSDYMNMTPGACAGGCGTGCCCTCCCGGTTTAGTGGTA RRPGPTRKHYQPYAPPGCGGAAGCGGTACGGATTTTACCTTTACT RDFAAYRSRVKFSRSAATCAGCTCTCTCCAACCCGAAGACATTGC DAPAYQQGQNQLYNELAACCTACTATTGTCAACAATATGGAAACC NLGRREEYDVLDKRRGTGCCTTTTACATTTGGCGGCGGCACCAAG RDPEMGGKPRRKNPQEGTGGAGATTAAGCGGGCGGCAGCTATTGA GLYNELQKDKMAEAYSGGTGATGTATCCACCGCCTTACCTGGATA EIGMKGERRRGKGHDGACGAAAAGAGTAACGGTACCATCATTCAC LYQGLSTATKDTYDALGTGAAAGGTAAACACCTGTGTCCTTCTCC HMQALPPR CCTCTTCCCCGGGCCATCAAAGCCCTTCTGGGTTCTTGTGGTCGTGGGAGGCGTGCTT GCTTGTTATTCTCTGCTCGTTACCGTGGCGTTTATCATTTTTTGGGTTAGATCCAAAA GAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCAC AAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGG GTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGT ATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGG ACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTAT AATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAG AGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAA GGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR1.2) CAGGTGCAGCTGCAGGAATCCGGACCGGG 171QVQLQESGPGLVKPSE 172 Clone 24C1 GCTGGTGAAGCCCAGCGAGACTCTGAGTCTLSLTCTVSGGSISSY CHD CAR TCACGTGTACAGTTTCTGGAGGTAGCATT YWSWIRQPPGKGLEWIDNA HxL AGCTCCTACTATTGGTCATGGATAAGGCA GYIYYSGSTNYNPSLKGCCCCCCGGGAAGGGATTGGAATGGATCG SRVTISVDTSKNQFSLGCTATATTTACTACAGTGGGAGCACCAAT KLSSVTAADTAVYYCVTACAACCCCTCACTGAAGTCTAGAGTTAC SLVYCGGDCYSGFDYWAATCAGCGTTGACACCTCAAAGAATCAGT GQGTLVTVSSGGGGSGTCAGTTTGAAATTGTCTAGCGTCACAGCA GGGSGGGGSDIQLTQSGCTGATACAGCCGTCTATTATTGTGTTTC PSSLSASVGDRVSFTCTCTGGTCTATTGCGGTGGGGATTGTTACA QASQDINNFLNWYQQKGTGGCTTTGACTATTGGGGGCAGGGTACT PGKAPKLLIYDASNLECTGGTTACAGTTTCTTCCGGGGGGGGAGG TGVPSRFSGSGSGTDFCTCTGGGGGCGGAGGCTCAGGTGGTGGAG TFTISSLQPEDIATYYGCAGCGACATCCAGTTGACACAGAGCCCG CQQYGNLPFTFGGGTKAGTTCCTTGTCCGCCTCCGTCGGGGATAG VEIKRAAAIEVMYPPPAGTGTCATTTACCTGTCAGGCCTCTCAGG YLDNEKSNGTIIHVKGATATTAATAACTTTCTGAATTGGTATCAG KHLCPSPLFPGPSKPFCAAAAGCCCGGAAAGGCACCCAAGCTGTT WVLVVVGGVLACYSLLGATTTACGACGCCAGTAACCTGGAGACAG VTVAFIIFWVRSKRSRGCGTGCCCTCCCGGTTTAGTGGTAGCGGA LLHSDYMNMTPRRPGPAGCGGTACGGATTTTACCTTTACTATCAG TRKHYQPYAPPRDFAACTCTCTCCAACCCGAAGACATTGCAACCT YRSRVKFSRSADAPAYACTATTGTCAACAATATGGAAACCTGCCT QQGQNQLYNELNLGRRTTTACATTTGGCGGCGGCACCAAGGTGGA EEYDVLDKRRGRDPEMGATTAAGCGGGCGGCAGCTATTGAGGTGA GGKPRRKNPQEGLYNETGTATCCACCGCCTTACCTGGATAACGAA LQKDKMAEAYSEIGMKAAGAGTAACGGTACCATCATTCACGTGAA GERRRGKGHDGLYQGLAGGTAAACACCTGTGTCCTTCTCCCCTCT STATKDTYDALHMQALTCCCCGGGCCATCAAAGCCCTTCTGGGTT PPR CTTGTGGTCGTGGGAGGCGTGCTTGCTTGTTATTCTCTGCTCGTTACCGTGGCGTTTA TCATTTTTTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATAT GACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGA GATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGT ATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTA TGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGA AAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCT ATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTA CCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTG CCACCTAGG (CAR1.3)ATGGCACTCCCCGTAACTGCTCTGCTGCT 173 MALPVTALLLPLALLL 174 Clone 24C1GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL CD8 CARGCCCGCAGGTGCAATTGCAAGAGTCCGGC VKPSETLSLTCTVSGG DNA HxLCCCGGACTCGTTAAACCCAGTGAGACGCT SISSYYWSWIRQPPGKTAGCCTGACCTGTACCGTCTCAGGGGGCA GLEWIGYIYYSGSTNYGCATCTCCTCTTATTACTGGAGCTGGATC NPSLKSRVTISVDTSKAGGCAGCCTCCAGGAAAAGGCCTTGAATG NQFSLKLSSVTAADTAGATTGGGTACATCTACTACTCTGGCTCAA VYYCVSLVYCGGDCYSCAAATTATAATCCATCCCTGAAGTCCCGC GFDYWGQGTLVTVSSGGTGACTATCTCTGTGGACACCAGCAAGAA GGGSGGGGSGGGGSDITCAGTTTTCACTGAAGTTGTCTAGTGTTA QLTQSPSSLSASVGDRCCGCGGCCGACACCGCCGTATACTACTGT VSFTCQASQDINNFLNGTGTCTCTTGTGTACTGTGGCGGCGACTG WYQQKPGKAPKLLIYDCTATTCCGGGTTCGACTACTGGGGCCAAG ASNLETGVPSRFSGSGGGACTCTGGTAACCGTGTCCTCAGGCGGC SGTDFTFTISSLQPEDGGCGGGTCAGGAGGAGGCGGCAGTGGAGG IATYYCQQYGNLPFTFTGGCGGCTCCGACATCCAGCTGACACAAT GGGTKVEIKRAAALSNCACCATCTTCCCTTTCAGCTTCAGTCGGG SIMYFSHFVPVFLPAKGACAGAGTGTCCTTCACATGCCAGGCCAG PTTTPAPRPPTPAPTICCAGGATATCAATAACTTCCTGAACTGGT ASQPLSLRPEACRPAAACCAACAGAAACCCGGAAAGGCTCCAAAG GGAVHTRGLDFACDIYCTCCTGATCTATGATGCTTCCAACCTGGA IWAPLAGTCGVLLLSLGACCGGCGTGCCCTCCAGGTTCAGTGGTT VITLYCNHRNRSKRSRCAGGATCAGGCACTGACTTTACGTTCACC LLHSDYMNMTPRRPGPATATCCAGTCTTCAGCCCGAAGACATTGC TRKHYQPYAPPRDFAAAACCTATTACTGCCAACAATACGGGAACC YRSRVKFSRSADAPAYTTCCCTTTACATTCGGAGGCGGCACCAAG QQGQNQLYNELNLGRRGTGGAAATCAAAAGGGCTGCAGCATTGAG EEYDVLDKRRGRDPEMCAACTCAATAATGTATTTTAGTCACTTTG GGKPRRKNPQEGLYNETACCAGTGTTCTTGCCGGCTAAGCCTACT LQKDKMAEAYSEIGMKACCACACCCGCTCCACGGCCACCTACCCC GERRRGKGHDGLYQGLAGCTCCTACCATCGCTTCACAGCCTCTGT STATKDTYDALHMQALCCCTGCGCCCAGAGGCTTGCCGACCGGCC PPR GCAGGGGGCGCTGTTCATACCAGAGGACTGGATTTCGCCTGCGATATCTATATCTGGG CACCCCTGGCCGGAACCTGCGGCGTACTCCTGCTGTCCCTGGTCATCACGCTCTATTG TAATCACAGGAACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATG ACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAG ATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTA TCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTAT GACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAA AAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTA TTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTAC CAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGC CACCTAGGTAA (CAR1.3)CAGGTGCAATTGCAAGAGTCCGGCCCCGG 175 QVQLQESGPGLVKPSE 176 Clone 24C1ACTCGTTAAACCCAGTGAGACGCTTAGCC TLSLTCTVSGGSISSY CD8 CARTGACCTGTACCGTCTCAGGGGGCAGCATC YWSWIRQPPGKGLEWI DNA HxLTCCTCTTATTACTGGAGCTGGATCAGGCA GYIYYSGSTNYNPSLKGCCTCCAGGAAAAGGCCTTGAATGGATTG SRVTISVDTSKNQFSLGGTACATCTACTACTCTGGCTCAACAAAT KLSSVTAADTAVYYCVTATAATCCATCCCTGAAGTCCCGCGTGAC SLVYCGGDCYSGFDYWTATCTCTGTGGACACCAGCAAGAATCAGT GQGTLVTVSSGGGGSGTTTCACTGAAGTTGTCTAGTGTTACCGCG GGGSGGGGSDIQLTQSGCCGACACCGCCGTATACTACTGTGTGTC PSSLSASVGDRVSFTCTCTTGTGTACTGTGGCGGCGACTGCTATT QASQDINNFLNWYQQKCCGGGTTCGACTACTGGGGCCAAGGGACT PGKAPKLLIYDASNLECTGGTAACCGTGTCCTCAGGCGGCGGCGG TGVPSRFSGSGSGTDFGTCAGGAGGAGGCGGCAGTGGAGGTGGCG TFTISSLQPEDIATYYGCTCCGACATCCAGCTGACACAATCACCA CQQYGNLPFTFGGGTKTCTTCCCTTTCAGCTTCAGTCGGGGACAG VEIKRAAALSNSIMYFAGTGTCCTTCACATGCCAGGCCAGCCAGG SHFVPVFLPAKPTTTPATATCAATAACTTCCTGAACTGGTACCAA APRPPTPAPTIASQPLCAGAAACCCGGAAAGGCTCCAAAGCTCCT SLRPEACRPAAGGAVHGATCTATGATGCTTCCAACCTGGAGACCG TRGLDFACDIYIWAPLGCGTGCCCTCCAGGTTCAGTGGTTCAGGA AGTCGVLLLSLVITLYTCAGGCACTGACTTTACGTTCACCATATC CNHRNRSKRSRLLHSDCAGTCTTCAGCCCGAAGACATTGCAACCT YMNMTPRRPGPTRKHYATTACTGCCAACAATACGGGAACCTTCCC QPYAPPRDFAAYRSRVTTTACATTCGGAGGCGGCACCAAGGTGGA KFSRSADAPAYQQGQNAATCAAAAGGGCTGCAGCATTGAGCAACT QLYNELNLGRREEYDVCAATAATGTATTTTAGTCACTTTGTACCA LDKRRGRDPEMGGKPRGTGTTCTTGCCGGCTAAGCCTACTACCAC RKNPQEGLYNELQKDKACCCGCTCCACGGCCACCTACCCCAGCTC MAEAYSEIGMKGERRRCTACCATCGCTTCACAGCCTCTGTCCCTG GKGHDGLYQGLSTATKCGCCCAGAGGCTTGCCGACCGGCCGCAGG DTYDALHMQALPPRGGGCGCTGTTCATACCAGAGGACTGGATT TCGCCTGCGATATCTATATCTGGGCACCCCTGGCCGGAACCTGCGGCGTACTCCTGCT GTCCCTGGTCATCACGCTCTATTGTAATCACAGGAACAGATCCAAAAGAAGCCGCCTG CTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACT ACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTC CAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTC AACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTG AGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCA GAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGG GGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATG ACGCTCTCCACATGCAAGCCCTGCCACCT AGG (CAR1.4)ATGGCACTCCCCGTAACTGCTCTGCTGCT 177 MALPVTALLLPLALLL 178 Clone 24C1GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPDIQLTQSPSSL THD CARGCCCGGATATCCAGCTCACGCAATCCCCC SASVGDRVSFTCQASQ DNA LxHTCAAGCTTGAGTGCCTCCGTGGGCGACCG DINNFLNWYQQKPGKAGGTGTCCTTCACATGTCAGGCAAGCCAAG PKLLIYDASNLETGVPACATAAATAATTTCCTGAATTGGTACCAA SRFSGSGSGTDFTFTICAAAAACCCGGCAAGGCTCCCAAACTCCT SSLQPEDIATYYCQQYGATTTATGATGCCTCCAATCTGGAGACCG GNLPFTFGGGTKVEIKGGGTCCCTTCTAGATTCAGCGGAAGTGGC RGGGGSGGGGSGGGGSAGCGGCACAGACTTTACATTTACTATCTC QVQLQESGPGLVKPSETTCTCTGCAACCAGAGGACATCGCCACAT TLSLTCTVSGGSISSYACTATTGCCAGCAATACGGCAATCTGCCC YWSWIRQPPGKGLEWITTCACCTTCGGAGGCGGAACCAAGGTAGA GYIYYSGSTNYNPSLKAATTAAAAGGGGCGGTGGAGGCTCCGGAG SRVTISVDTSKNQFSLGGGGGGGCTCTGGCGGAGGGGGCTCCCAA KLSSVTAADTAVYYCVGTACAATTGCAGGAGTCAGGGCCTGGACT SLVYCGGDCYSGFDYWCGTGAAGCCTTCAGAAACTTTGTCACTGA GQGTLVTVSSAAALDNCATGTACAGTGTCCGGCGGAAGCATTTCC EKSNGTIIHVKGKHLCAGTTACTATTGGTCCTGGATTAGACAGCC PSPLFPGPSKPFWVLVACCCGGCAAAGGACTGGAATGGATTGGAT VVGGVLACYSLLVTVAATATCTACTACTCTGGATCTACAAACTAT FIIFWVRSKRSRLLHSAATCCCAGCCTCAAATCCAGGGTCACTAT DYMNMTPRRPGPTRKHTAGTGTGGATACATCAAAGAATCAGTTCT YQPYAPPRDFAAYRSRCCTTGAAGCTGAGCTCAGTCACTGCTGCC VKFSRSADAPAYQQGQGACACCGCAGTGTACTATTGTGTGAGCCT NQLYNELNLGRREEYDGGTCTACTGCGGCGGAGATTGCTACAGCG VLDKRRGRDPEMGGKPGTTTCGATTACTGGGGCCAGGGCACCCTG RRKNPQEGLYNELQKDGTTACCGTTAGTTCCGCGGCTGCTCTTGA KMAEAYSEIGMKGERRTAACGAGAAGTCCAACGGTACGATTATCC RGKGHDGLYQGLSTATACGTTAAGGGTAAGCACCTTTGCCCTAGC KDTYDALHMQALPPRCCGCTGTTCCCAGGCCCCAGTAAGCCCTT TTGGGTCCTCGTTGTGGTAGGTGGGGTACTCGCCTGCTACTCCCTGCTCGTCACTGTC GCATTCATCATCTTCTGGGTCAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACA TGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACC ACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCA CCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGG AAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACC AAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCT GAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACG GTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCA AGCCCTGCCACCTAGGTAA (CAR1.4)GATATCCAGCTCACGCAATCCCCCTCAAG 179 DIQLTQSPSSLSASVG 180 Clone 24C1CTTGAGTGCCTCCGTGGGCGACCGGGTGT DRVSFTCQASQDINNF THD CARCCTTCACATGTCAGGCAAGCCAAGACATA LNWYQQKPGKAPKLLI DNA LxHAATAATTTCCTGAATTGGTACCAACAAAA YDASNLETGVPSRFSGACCCGGCAAGGCTCCCAAACTCCTGATTT SGSGTDFTFTISSLQPATGATGCCTCCAATCTGGAGACCGGGGTC EDIATYYCQQYGNLPFCCTTCTAGATTCAGCGGAAGTGGCAGCGG TFGGGTKVEIKRGGGGCACAGACTTTACATTTACTATCTCTTCTC SGGGGSGGGGSQVQLQTGCAACCAGAGGACATCGCCACATACTAT ESGPGLVKPSETLSLTTGCCAGCAATACGGCAATCTGCCCTTCAC CTVSGGSISSYYWSWICTTCGGAGGCGGAACCAAGGTAGAAATTA RQPPGKGLEWIGYIYYAAAGGGGCGGTGGAGGCTCCGGAGGGGGG SGSTNYNPSLKSRVTIGGCTCTGGCGGAGGGGGCTCCCAAGTACA SVDTSKNQFSLKLSSVATTGCAGGAGTCAGGGCCTGGACTCGTGA TAADTAVYYCVSLVYCAGCCTTCAGAAACTTTGTCACTGACATGT GGDCYSGFDYWGQGTLACAGTGTCCGGCGGAAGCATTTCCAGTTA VTVSSAAALDNEKSNGCTATTGGTCCTGGATTAGACAGCCACCCG TIIHVKGKHLCPSPLFGCAAAGGACTGGAATGGATTGGATATATC PGPSKPFWVLVVVGGVTACTACTCTGGATCTACAAACTATAATCC LACYSLLVTVAFIIFWCAGCCTCAAATCCAGGGTCACTATTAGTG VRSKRSRLLHSDYMNMTGGATACATCAAAGAATCAGTTCTCCTTG TPRRPGPTRKHYQPYAAAGCTGAGCTCAGTCACTGCTGCCGACAC PPRDFAAYRSRVKFSRCGCAGTGTACTATTGTGTGAGCCTGGTCT SADAPAYQQGQNQLYNACTGCGGCGGAGATTGCTACAGCGGTTTC ELNLGRREEYDVLDKRGATTACTGGGGCCAGGGCACCCTGGTTAC RGRDPEMGGKPRRKNPCGTTAGTTCCGCGGCTGCTCTTGATAACG QEGLYNELQKDKMAEAAGAAGTCCAACGGTACGATTATCCACGTT YSEIGMKGERRRGKGHAAGGGTAAGCACCTTTGCCCTAGCCCGCT DGLYQGLSTATKDTYDGTTCCCAGGCCCCAGTAAGCCCTTTTGGG ALHMQALPPR TCCTCGTTGTGGTAGGTGGGGTACTCGCCTGCTACTCCCTGCTCGTCACTGTCGCATT CATCATCTTCTGGGTCAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAAT ATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTA GAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGC GTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAG TATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGAC GAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGC CTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTG TACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCC TGCCACCTAGG (CAR1.5)ATGGCACTCCCCGTAACTGCTCTGCTGCT 181 MALPVTALLLPLALLL 182 Clone 24C1GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPDIQLTQSPSSL CHD CARGCCCGGATATCCAGCTGACCCAGTCTCCA SASVGDRVSFTCQASQ DNA LxHTCCTCTTTGAGTGCCTCCGTGGGTGACCG DINNFLNWYQQKPGKACGTCTCTTTCACTTGCCAAGCCAGCCAAG PKLLIYDASNLETGVPACATCAACAACTTTCTGAATTGGTACCAG SRFSGSGSGTDFTFTICAGAAACCAGGCAAAGCACCAAAGCTCCT SSLQPEDIATYYCQQYCATCTACGACGCCTCCAACCTGGAAACCG GNLPFTFGGGTKVEIKGGGTGCCCAGCAGGTTTAGCGGGAGCGGT RGGGGSGGGGSGGGGSTCTGGCACGGATTTTACGTTCACCATCTC QVQLQESGPGLVKPSECTCTCTGCAGCCCGAGGATATAGCTACTT TLSLTCTVSGGSISSYATTACTGTCAGCAGTACGGGAATCTGCCA YWSWIRQPPGKGLEWITTTACTTTTGGGGGTGGAACTAAGGTGGA GYIYYSGSTNYNPSLKAATCAAAAGGGGCGGCGGGGGAAGCGGGG SRVTISVDTSKNQFSLGCGGGGGCTCAGGTGGCGGAGGGAGCCAG KLSSVTAADTAVYYCVGTGCAACTCCAGGAAAGTGGCCCAGGATT SLVYCGGDCYSGFDYWGGTGAAGCCCAGCGAGACCCTTTCCCTTA GQGTLVTVSSAAAIEVCTTGTACTGTTAGCGGAGGCAGCATAAGC MYPPPYLDNEKSNGTIAGCTACTATTGGTCCTGGATCAGACAGCC IHVKGKHLCPSPLFPGACCAGGGAAAGGGCTTGAATGGATTGGCT PSKPFWVLVVVGGVLAACATTTACTATTCCGGGTCCACCAACTAC CYSLLVTVAFIIFWVRAACCCATCCCTCAAGTCCCGCGTGACAAT SKRSRLLHSDYMNMTPTTCCGTCGACACAAGCAAGAACCAGTTCT RRPGPTRKHYQPYAPPCCCTGAAACTTAGTAGCGTCACTGCTGCA RDFAAYRSRVKFSRSAGATACAGCAGTGTACTATTGTGTCAGCCT DAPAYQQGQNQLYNELTGTCTACTGTGGCGGCGACTGCTACAGTG NLGRREEYDVLDKRRGGCTTTGATTACTGGGGACAGGGCACGCTC RDPEMGGKPRRKNPQEGTGACAGTGTCCAGCGCTGCGGCTATCGA GLYNELQKDKMAEAYSGGTAATGTATCCGCCACCGTATCTGGACA EIGMKGERRRGKGHDGACGAGAAGTCTAATGGGACAATCATTCAC LYQGLSTATKDTYDALGTGAAGGGGAAGCACCTGTGTCCATCCCC HMQALPPR CCTGTTTCCGGGTCCCAGTAAACCCTTCTGGGTGCTTGTTGTCGTTGGCGGGGTGCTG GCCTGCTATTCCCTGCTGGTGACCGTCGCGTTTATTATTTTCTGGGTTAGATCCAAAA GAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCAC AAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGG GTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGT ATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGG ACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTAT AATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAG AGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAA GGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR1.5) GATATCCAGCTGACCCAGTCTCCATCCTC 183DIQLTQSPSSLSASVG 184 Clone 24C1 TTTGAGTGCCTCCGTGGGTGACCGCGTCTDRVSFTCQASQDINNF CHD CAR CTTTCACTTGCCAAGCCAGCCAAGACATC LNWYQQKPGKAPKLLIDNA LxH AACAACTTTCTGAATTGGTACCAGCAGAA YDASNLETGVPSRFSGACCAGGCAAAGCACCAAAGCTCCTCATCT SGSGTDFTFTISSLQPACGACGCCTCCAACCTGGAAACCGGGGTG EDIATYYCQQYGNLPFCCCAGCAGGTTTAGCGGGAGCGGTTCTGG TFGGGTKVEIKRGGGGCACGGATTTTACGTTCACCATCTCCTCTC SGGGGSGGGGSQVQLQTGCAGCCCGAGGATATAGCTACTTATTAC ESGPGLVKPSETLSLTTGTCAGCAGTACGGGAATCTGCCATTTAC CTVSGGSISSYYWSWITTTTGGGGGTGGAACTAAGGTGGAAATCA RQPPGKGLEWIGYIYYAAAGGGGCGGCGGGGGAAGCGGGGGCGGG SGSTNYNPSLKSRVTIGGCTCAGGTGGCGGAGGGAGCCAGGTGCA SVDTSKNQFSLKLSSVACTCCAGGAAAGTGGCCCAGGATTGGTGA TAADTAVYYCVSLVYCAGCCCAGCGAGACCCTTTCCCTTACTTGT GGDCYSGFDYWGQGTLACTGTTAGCGGAGGCAGCATAAGCAGCTA VTVSSAAAIEVMYPPPCTATTGGTCCTGGATCAGACAGCCACCAG YLDNEKSNGTIIHVKGGGAAAGGGCTTGAATGGATTGGCTACATT KHLCPSPLFPGPSKPFTACTATTCCGGGTCCACCAACTACAACCC WVLVVVGGVLACYSLLATCCCTCAAGTCCCGCGTGACAATTTCCG VTVAFIIFWVRSKRSRTCGACACAAGCAAGAACCAGTTCTCCCTG LLHSDYMNMTPRRPGPAAACTTAGTAGCGTCACTGCTGCAGATAC TRKHYQPYAPPRDFAAAGCAGTGTACTATTGTGTCAGCCTTGTCT YRSRVKFSRSADAPAYACTGTGGCGGCGACTGCTACAGTGGCTTT QQGQNQLYNELNLGRRGATTACTGGGGACAGGGCACGCTCGTGAC EEYDVLDKRRGRDPEMAGTGTCCAGCGCTGCGGCTATCGAGGTAA GGKPRRKNPQEGLYNETGTATCCGCCACCGTATCTGGACAACGAG LQKDKMAEAYSEIGMKAAGTCTAATGGGACAATCATTCACGTGAA GERRRGKGHDGLYQGLGGGGAAGCACCTGTGTCCATCCCCCCTGT STATKDTYDALHMQALTTCCGGGTCCCAGTAAACCCTTCTGGGTG PPR CTTGTTGTCGTTGGCGGGGTGCTGGCCTGCTATTCCCTGCTGGTGACCGTCGCGTTTA TTATTTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATAT GACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGA GATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGT ATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTA TGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGA AAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCT ATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTA CCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTG CCACCTAGG (CAR1.6)ATGGCACTCCCCGTAACTGCTCTGCTGCT 185 MALPVTALLLPLALLL 186 Clone 24C1GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPDIQLTQSPSSL CD8 CARGCCCGGACATTCAATTGACCCAGTCCCCT SASVGDRVSFTCQASQ DNA LxHAGCAGTCTCTCAGCAAGTGTGGGAGATAG DINNFLNWYQQKPGKAGGTGTCATTCACCTGTCAGGCTTCACAGG PKLLIYDASNLETGVPACATCAACAACTTCCTCAATTGGTATCAG SRFSGSGSGTDFTFTICAGAAGCCAGGGAAGGCACCAAAGCTGCT SSLQPEDIATYYCQQYCATATATGACGCTTCAAACCTTGAAACCG GNLPFTFGGGTKVEIKGAGTACCTAGCCGCTTCAGCGGAAGCGGA RGGGGSGGGGSGGGGSTCAGGGACTGACTTCACTTTTACCATCTC QVQLQESGPGLVKPSETTCACTGCAGCCCGAAGACATCGCCACAT TLSLTCTVSGGSISSYACTACTGCCAGCAGTACGGAAACTTGCCT YWSWIRQPPGKGLEWITTTACATTTGGGGGCGGCACCAAAGTGGA GYIYYSGSTNYNPSLKGATTAAGCGAGGGGGAGGCGGCTCAGGAG SRVTISVDTSKNQFSLGCGGTGGCTCCGGAGGCGGGGGTTCCCAG KLSSVTAADTAVYYCVGTCCAGCTCCAGGAATCCGGCCCAGGTCT SLVYCGGDCYSGFDYWGGTTAAGCCCAGTGAAACTTTGTCCCTCA GQGTLVTVSSAAALSNCGTGTACTGTGAGCGGTGGTTCAATCTCC SIMYFSHFVPVFLPAKTCATACTATTGGTCTTGGATACGGCAACC PTTTPAPRPPTPAPTITCCTGGAAAGGGCCTCGAGTGGATCGGCT ASQPLSLRPEACRPAAATATCTACTATAGTGGCTCCACTAATTAC GGAVHTRGLDFACDIYAACCCTTCCCTCAAGTCCAGAGTCACCAT IWAPLAGTCGVLLLSLTTCCGTGGACACATCTAAGAACCAGTTCA VITLYCNHRNRSKRSRGTCTGAAGTTGTCCAGCGTTACAGCCGCA LLHSDYMNMTPRRPGPGACACAGCCGTTTATTACTGTGTGTCTCT TRKHYQPYAPPRDFAATGTTTACTGCGGGGGAGACTGTTATAGCG YRSRVKFSRSADAPAYGCTTCGATTACTGGGGCCAGGGCACCTTG QQGQNQLYNELNLGRRGTCACAGTCTCTTCCGCGGCCGCCCTCTC EEYDVLDKRRGRDPEMTAACAGTATTATGTACTTTTCTCATTTTG GGKPRRKNPQEGLYNETACCCGTGTTCCTTCCCGCTAAGCCAACT LQKDKMAEAYSEIGMKACTACCCCGGCCCCACGGCCGCCTACCCC GERRRGKGHDGLYQGLTGCACCCACAATAGCCAGTCAGCCTTTGA STATKDTYDALHMQALGCCTGAGACCTGAGGCTTGTCGGCCGGCT PPR GCTGGGGGTGCAGTGCACACACGAGGTCTTGATTTTGCTTGCGACATATACATCTGGG CCCCTCTGGCCGGGACCTGTGGGGTGCTGCTTCTGAGCTTGGTCATCACGCTCTATTG CAACCATCGCAACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATG ACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAG ATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTA TCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTAT GACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAA AAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTA TTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTAC CAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGC CACCTAGGTAA (CAR1.6)GACATTCAATTGACCCAGTCCCCTAGCAG 187 DIQLTQSPSSLSASVG 188 Clone 24C1TCTCTCAGCAAGTGTGGGAGATAGGGTGT DRVSFTCQASQDINNF CD8 CARCATTCACCTGTCAGGCTTCACAGGACATC LNWYQQKPGKAPKLLI DNA LxHAACAACTTCCTCAATTGGTATCAGCAGAA YDASNLETGVPSRFSGGCCAGGGAAGGCACCAAAGCTGCTCATAT SGSGTDFTFTISSLQPATGACGCTTCAAACCTTGAAACCGGAGTA EDIATYYCQQYGNLPFCCTAGCCGCTTCAGCGGAAGCGGATCAGG TFGGGTKVEIKRGGGGGACTGACTTCACTTTTACCATCTCTTCAC SGGGGSGGGGSQVQLQTGCAGCCCGAAGACATCGCCACATACTAC ESGPGLVKPSETLSLTTGCCAGCAGTACGGAAACTTGCCTTTTAC CTVSGGSISSYYWSWIATTTGGGGGCGGCACCAAAGTGGAGATTA RQPPGKGLEWIGYIYYAGCGAGGGGGAGGCGGCTCAGGAGGCGGT SGSTNYNPSLKSRVTIGGCTCCGGAGGCGGGGGTTCCCAGGTCCA SVDTSKNQFSLKLSSVGCTCCAGGAATCCGGCCCAGGTCTGGTTA TAADTAVYYCVSLVYCAGCCCAGTGAAACTTTGTCCCTCACGTGT GGDCYSGFDYWGQGTLACTGTGAGCGGTGGTTCAATCTCCTCATA VTVSSAAALSNSIMYFCTATTGGTCTTGGATACGGCAACCTCCTG SHFVPVFLPAKPTTTPGAAAGGGCCTCGAGTGGATCGGCTATATC APRPPTPAPTIASQPLTACTATAGTGGCTCCACTAATTACAACCC SLRPEACRPAAGGAVHTTCCCTCAAGTCCAGAGTCACCATTTCCG TRGLDFACDIYIWAPLTGGACACATCTAAGAACCAGTTCAGTCTG AGTCGVLLLSLVITLYAAGTTGTCCAGCGTTACAGCCGCAGACAC CNHRNRSKRSRLLHSDAGCCGTTTATTACTGTGTGTCTCTTGTTT YMNMTPRRPGPTRKHYACTGCGGGGGAGACTGTTATAGCGGCTTC QPYAPPRDFAAYRSRVGATTACTGGGGCCAGGGCACCTTGGTCAC KFSRSADAPAYQQGQNAGTCTCTTCCGCGGCCGCCCTCTCTAACA QLYNELNLGRREEYDVGTATTATGTACTTTTCTCATTTTGTACCC LDKRRGRDPEMGGKPRGTGTTCCTTCCCGCTAAGCCAACTACTAC RKNPQEGLYNELQKDKCCCGGCCCCACGGCCGCCTACCCCTGCAC MAEAYSEIGMKGERRRCCACAATAGCCAGTCAGCCTTTGAGCCTG GKGHDGLYQGLSTATKAGACCTGAGGCTTGTCGGCCGGCTGCTGG DTYDALHMQALPPRGGGTGCAGTGCACACACGAGGTCTTGATT TTGCTTGCGACATATACATCTGGGCCCCTCTGGCCGGGACCTGTGGGGTGCTGCTTCT GAGCTTGGTCATCACGCTCTATTGCAACCATCGCAACAGATCCAAAAGAAGCCGCCTG CTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACT ACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTC CAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTC AACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTG AGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCA GAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGG GGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATG ACGCTCTCCACATGCAAGCCCTGCCACCT AGG (CAR2.1)ATGGCACTCCCCGTAACTGCTCTGCTGCT 189 MALPVTALLLPLALLL 190 Clone 24C8GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL THD CARGCCCGCAGGTACAGCTGCAGGAATCTGGG VKPSQTLSLTCTVSGG DNA HxLCCCGGACTTGTCAAGCCAAGTCAGACACT SISSGGFYWSWIRQHPTTCTCTTACATGTACCGTGAGCGGCGGAA GKGLEWIGYIHHSGSTGTATAAGCAGTGGAGGCTTTTACTGGTCT HYNPSLKSRVTISIDTTGGATACGGCAGCACCCAGGCAAAGGCTT SKNLFSLRLSSVTAADGGAGTGGATTGGATACATTCATCATTCAG TAVYYCASLVYCGGDCGATCTACACACTATAATCCATCCCTTAAG YSGFDYWGQGTLVTVSTCCCGGGTCACCATTAGCATTGATACGTC SGGGGSGGGGSGGGGSTAAGAATCTGTTCAGTCTCAGGCTGTCCT DIQLTQSPSSLSASVGCCGTCACTGCTGCCGACACAGCCGTGTAC DRVSFTCQASQDINNFTACTGCGCCTCCTTGGTTTACTGCGGAGG LNWYQQKPGKAPKLLICGACTGTTATAGCGGCTTTGATTATTGGG YDASNLETGVPSRFSGGGCAGGGGACCCTCGTAACCGTGAGCTCT SGSGTDFTFTISSLQPGGAGGGGGTGGGAGCGGGGGAGGAGGTTC EDIATYYCQQYGNLPFAGGGGGGGGCGGCTCCGATATCCAGCTCA TFGGGTKVEIKRAAALCTCAAAGCCCCTCTAGTCTCTCTGCCTCA DNEKSNGTIIHVKGKHGTGGGGGATCGGGTCAGTTTTACTTGTCA LCPSPLFPGPSKPFWVAGCTTCACAGGATATCAACAACTTCCTTA LVVVGGVLACYSLLVTATTGGTATCAGCAGAAGCCAGGAAAAGCA VAFIIFWVRSKRSRLLCCCAAGCTGCTCATCTATGATGCCTCAAA HSDYMNMTPRRPGPTRTTTGGAGACGGGTGTTCCCAGTCGATTCT KHYQPYAPPRDFAAYRCTGGGTCAGGGTCCGGGACCGACTTTACG SRVKFSRSADAPAYQQTTTACGATCTCCTCTCTGCAGCCCGAAGA GQNQLYNELNLGRREECATCGCCACATACTATTGTCAACAGTACG YDVLDKRRGRDPEMGGGCAACTTGCCTTTCACATTTGGGGGCGGG KPRRKNPQEGLYNELQACTAAGGTTGAAATCAAGAGGGCCGCTGC KDKMAEAYSEIGMKGEACTGGACAATGAGAAGTCCAACGGCACCA RRRGKGHDGLYQGLSTTCATCCACGTGAAGGGCAAGCACCTGTGC ATKDTYDALHMQALPPCCTAGTCCTCTGTTCCCAGGCCCATCCAA R ACCTTTTTGGGTTCTTGTTGTGGTCGGGGGGGTGCTGGCCTGCTATTCTCTGCTGGTC ACGGTGGCCTTCATAATTTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCG ATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTA CGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCA GATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGAC GCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGG CAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAG ATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGC ACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCA CATGCAAGCCCTGCCACCTAGGTAA (CAR2.1)CAGGTACAGCTGCAGGAATCTGGGCCCGG 191 QVQLQESGPGLVKPSQ 192 Clone 24C8ACTTGTCAAGCCAAGTCAGACACTTTCTC TLSLTCTVSGGSISSG THD CARTTACATGTACCGTGAGCGGCGGAAGTATA GFYWSWIRQHPGKGLE DNA HxLAGCAGTGGAGGCTTTTACTGGTCTTGGAT WIGYIHHSGSTHYNPSACGGCAGCACCCAGGCAAAGGCTTGGAGT LKSRVTISIDTSKNLFGGATTGGATACATTCATCATTCAGGATCT SLRLSSVTAADTAVYYACACACTATAATCCATCCCTTAAGTCCCG CASLVYCGGDCYSGFDGGTCACCATTAGCATTGATACGTCTAAGA YWGQGTLVTVSSGGGGATCTGTTCAGTCTCAGGCTGTCCTCCGTC SGGGGSGGGGSDIQLTACTGCTGCCGACACAGCCGTGTACTACTG QSPSSLSASVGDRVSFCGCCTCCTTGGTTTACTGCGGAGGCGACT TCQASQDINNFLNWYQGTTATAGCGGCTTTGATTATTGGGGGCAG QKPGKAPKLLIYDASNGGGACCCTCGTAACCGTGAGCTCTGGAGG LETGVPSRFSGSGSGTGGGTGGGAGCGGGGGAGGAGGTTCAGGGG DFTFTISSLQPEDIATGGGGCGGCTCCGATATCCAGCTCACTCAA YYCQQYGNLPFTFGGGAGCCCCTCTAGTCTCTCTGCCTCAGTGGG TKVEIKRAAALDNEKSGGATCGGGTCAGTTTTACTTGTCAAGCTT NGTIIHVKGKHLCPSPCACAGGATATCAACAACTTCCTTAATTGG LFPGPSKPFWVLVVVGTATCAGCAGAAGCCAGGAAAAGCACCCAA GVLACYSLLVTVAFIIGCTGCTCATCTATGATGCCTCAAATTTGG FWVRSKRSRLLHSDYMAGACGGGTGTTCCCAGTCGATTCTCTGGG NMTPRRPGPTRKHYQPTCAGGGTCCGGGACCGACTTTACGTTTAC YAPPRDFAAYRSRVKFGATCTCCTCTCTGCAGCCCGAAGACATCG SRSADAPAYQQGQNQLCCACATACTATTGTCAACAGTACGGCAAC YNELNLGRREEYDVLDTTGCCTTTCACATTTGGGGGCGGGACTAA KRRGRDPEMGGKPRRKGGTTGAAATCAAGAGGGCCGCTGCACTGG NPQEGLYNELQKDKMAACAATGAGAAGTCCAACGGCACCATCATC EAYSEIGMKGERRRGKCACGTGAAGGGCAAGCACCTGTGCCCTAG GHDGLYQGLSTATKDTTCCTCTGTTCCCAGGCCCATCCAAACCTT YDALHMQALPPR TTTGGGTTCTTGTTGTGGTCGGGGGGGTGCTGGCCTGCTATTCTCTGCTGGTCACGGT GGCCTTCATAATTTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTAC ATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCAC CACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGC ACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGG GAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAAC CAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGC TGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGAC GGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGC AAGCCCTGCCACCTAGG (CAR2.2)ATGGCACTCCCCGTAACTGCTCTGCTGCT 193 MALPVTALLLPLALLL 194 Clone 24C8GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL CHD CARGCCCGCAGGTGCAGCTGCAGGAAAGCGGT VKPSQTLSLTCTVSGG DNA HxLCCGGGACTTGTCAAGCCGTCCCAAACGCT SISSGGFYWSWIRQHPGAGTCTGACGTGTACTGTCTCTGGTGGCT GKGLEWIGYIHHSGSTCTATTTCTTCCGGGGGCTTTTATTGGTCT HYNPSLKSRVTISIDTTGGATCAGACAACACCCTGGCAAAGGGCT SKNLFSLRLSSVTAADGGAGTGGATAGGGTATATTCACCACTCTG TAVYYCASLVYCGGDCGGTCCACTCACTACAACCCATCATTGAAA YSGFDYWGQGTLVTVSTCCAGAGTGACTATCTCAATCGACACATC SGGGGSGGGGSGGGGSCAAGAACCTTTTCAGCCTGAGGTTGTCAT DIQLTQSPSSLSASVGCAGTTACCGCCGCTGACACCGCGGTGTAT DRVSFTCQASQDINNFTATTGCGCCTCTCTCGTGTACTGCGGTGG LNWYQQKPGKAPKLLICGATTGTTATAGTGGCTTTGACTACTGGG YDASNLETGVPSRFSGGGCAGGGGACATTGGTTACCGTTTCAAGT SGSGTDFTFTISSLQPGGAGGCGGTGGGTCTGGCGGGGGCGGTAG EDIATYYCQQYGNLPFCGGAGGTGGGGGGAGCGACATACAGCTTA TFGGGTKVEIKRAAAICGCAGAGCCCCTCCAGCCTTTCAGCCTCC EVMYPPPYLDNEKSNGGTGGGGGATAGGGTGTCCTTTACCTGCCA TIIHVKGKHLCPSPLFGGCTTCCCAGGACATAAACAACTTCCTCA PGPSKPFWVLVVVGGVATTGGTATCAGCAAAAGCCCGGGAAAGCA LACYSLLVTVAFIIFWCCAAAGCTGCTCATCTACGATGCCAGCAA VRSKRSRLLHSDYMNMCCTGGAAACCGGAGTGCCGTCTCGCTTCT TPRRPGPTRKHYQPYACTGGAAGTGGCAGTGGGACCGATTTCACT PPRDFAAYRSRVKFSRTTTACAATCTCAAGTTTGCAGCCAGAAGA SADAPAYQQGQNQLYNCATTGCAACATACTACTGTCAACAGTACG ELNLGRREEYDVLDKRGCAATCTCCCCTTTACATTTGGGGGGGGA RGRDPEMGGKPRRKNPACTAAAGTGGAGATTAAGCGCGCTGCAGC QEGLYNELQKDKMAEACATTGAAGTTATGTATCCGCCCCCGTATC YSEIGMKGERRRGKGHTGGATAACGAGAAATCTAATGGTACCATA DGLYQGLSTATKDTYDATACATGTGAAGGGGAAGCACCTCTGTCC ALHMQALPPR ATCACCGCTGTTCCCCGGCCCTTCAAAACCTTTCTGGGTACTCGTTGTCGTGGGTGGA GTTCTGGCCTGCTATAGTCTGCTGGTGACCGTGGCGTTTATCATCTTCTGGGTAAGAT CCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGG CCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGG AGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACC AACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCG CAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGT CTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGA AAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGC TACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR2.2) CAGGTGCAGCTGCAGGAAAGCGGTCCGGG 195QVQLQESGPGLVKPSQ 196 Clone 24C8 ACTTGTCAAGCCGTCCCAAACGCTGAGTCTLSLTCTVSGGSISSG CHD CAR TGACGTGTACTGTCTCTGGTGGCTCTATT GFYWSWIRQHPGKGLEDNA HxL TCTTCCGGGGGCTTTTATTGGTCTTGGAT WIGYIHHSGSTHYNPSCAGACAACACCCTGGCAAAGGGCTGGAGT LKSRVTISIDTSKNLFGGATAGGGTATATTCACCACTCTGGGTCC SLRLSSVTAADTAVYYACTCACTACAACCCATCATTGAAATCCAG CASLVYCGGDCYSGFDAGTGACTATCTCAATCGACACATCCAAGA YWGQGTLVTVSSGGGGACCTTTTCAGCCTGAGGTTGTCATCAGTT SGGGGSGGGGSDIQLTACCGCCGCTGACACCGCGGTGTATTATTG QSPSSLSASVGDRVSFCGCCTCTCTCGTGTACTGCGGTGGCGATT TCQASQDINNFLNWYQGTTATAGTGGCTTTGACTACTGGGGGCAG QKPGKAPKLLIYDASNGGGACATTGGTTACCGTTTCAAGTGGAGG LETGVPSRFSGSGSGTCGGTGGGTCTGGCGGGGGCGGTAGCGGAG DFTFTISSLQPEDIATGTGGGGGGAGCGACATACAGCTTACGCAG YYCQQYGNLPFTFGGGAGCCCCTCCAGCCTTTCAGCCTCCGTGGG TKVEIKRAAAIEVMYPGGATAGGGTGTCCTTTACCTGCCAGGCTT PPYLDNEKSNGTIIHVCCCAGGACATAAACAACTTCCTCAATTGG KGKHLCPSPLFPGPSKTATCAGCAAAAGCCCGGGAAAGCACCAAA PFWVLVVVGGVLACYSGCTGCTCATCTACGATGCCAGCAACCTGG LLVTVAFIIFWVRSKRAAACCGGAGTGCCGTCTCGCTTCTCTGGA SRLLHSDYMNMTPRRPAGTGGCAGTGGGACCGATTTCACTTTTAC GPTRKHYQPYAPPRDFAATCTCAAGTTTGCAGCCAGAAGACATTG AAYRSRVKFSRSADAPCAACATACTACTGTCAACAGTACGGCAAT AYQQGQNQLYNELNLGCTCCCCTTTACATTTGGGGGGGGAACTAA RREEYDVLDKRRGRDPAGTGGAGATTAAGCGCGCTGCAGCCATTG EMGGKPRRKNPQEGLYAAGTTATGTATCCGCCCCCGTATCTGGAT NELQKDKMAEAYSEIGAACGAGAAATCTAATGGTACCATAATACA MKGERRRGKGHDGLYQTGTGAAGGGGAAGCACCTCTGTCCATCAC GLSTATKDTYDALHMQCGCTGTTCCCCGGCCCTTCAAAACCTTTC ALPPR TGGGTACTCGTTGTCGTGGGTGGAGTTCTGGCCTGCTATAGTCTGCTGGTGACCGTGG CGTTTATCATCTTCTGGGTAAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACAT GAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCA CCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCAC CAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGA AGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCA AGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTG AAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGG TTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAA GCCCTGCCACCTAGG (CAR2.3)ATGGCACTCCCCGTAACTGCTCTGCTGCT 197 MALPVTALLLPLALLL 198 Clone 24C8GCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLQESGPGL CD8 CARGCCCGCAGGTGCAGTTGCAGGAAAGCGGG VKPSQTLSLTCTVSGG DNA HxLCCTGGCCTTGTGAAACCAAGCCAGACACT SISSGGFYWSWIRQHPGAGCCTGACATGCACTGTGTCCGGCGGGT GKGLEWIGYIHHSGSTCCATATCTTCCGGGGGTTTTTATTGGTCC HYNPSLKSRVTISIDTTGGATACGCCAGCATCCCGGGAAAGGACT SKNLFSLRLSSVTAADTGAATGGATTGGATATATCCACCATTCCG TAVYYCASLVYCGGDCGAAGCACCCACTACAATCCAAGCCTTAAA YSGFDYWGQGTLVTVSTCCCGGGTGACAATCTCCATCGACACCTC SGGGGSGGGGSGGGGSAAAGAATCTTTTTTCCCTGCGGTTGTCTT DIQLTQSPSSLSASVGCAGTAACTGCCGCCGATACCGCTGTGTAC DRVSFTCQASQDINNFTACTGTGCCAGCCTCGTCTATTGCGGCGG LNWYQQKPGKAPKLLIAGATTGTTATTCTGGGTTCGATTATTGGG YDASNLETGVPSRFSGGTCAAGGCACACTGGTAACTGTCAGCAGC SGSGTDFTFTISSLQPGGAGGCGGCGGTTCCGGGGGCGGGGGCAG EDIATYYCQQYGNLPFTGGAGGGGGCGGATCTGACATTCAGCTTA TFGGGTKVEIKRAAALCGCAGTCCCCATCTTCACTTAGCGCCAGC SNSIMYFSHFVPVFLPGTTGGCGATCGGGTCAGCTTCACGTGTCA AKPTTTPAPRPPTPAPAGCAAGTCAGGATATCAACAACTTTCTTA TIASQPLSLRPEACRPACTGGTACCAGCAGAAGCCAGGCAAGGCA AAGGAVHTRGLDFACDCCCAAGTTGCTGATTTACGATGCTTCTAA IYIWAPLAGTCGVLLLCCTCGAGACGGGAGTGCCTAGCCGCTTCT SLVITLYCNHRNRSKRCCGGGAGCGGCAGCGGCACAGACTTTACC SRLLHSDYMNMTPRRPTTTACGATTTCCAGTCTGCAGCCAGAGGA GPTRKHYQPYAPPRDFTATAGCAACTTATTACTGTCAGCAGTATG AAYRSRVKFSRSADAPGCAACCTCCCTTTTACCTTCGGTGGTGGC AYQQGQNQLYNELNLGACAAAGGTCGAGATTAAAAGAGCCGCAGC RREEYDVLDKRRGRDPGTTGTCCAACTCCATAATGTATTTTTCTC EMGGKPRRKNPQEGLYATTTTGTGCCCGTCTTTCTGCCTGCCAAA NELQKDKMAEAYSEIGCCTACCACCACCCCCGCCCCACGACCACC MKGERRRGKGHDGLYQTACTCCAGCCCCCACCATCGCCTCCCAGC GLSTATKDTYDALHMQCCCTCAGCCTGAGGCCAGAGGCTTGTCGC ALPPR CCTGCTGCGGGGGGCGCTGTCCATACCAGAGGACTCGACTTCGCCTGCGATATTTATA TATGGGCCCCCCTCGCCGGCACCTGCGGAGTCTTGCTCCTGAGCCTTGTGATCACGCT TTATTGTAACCATCGGAATAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATG AATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCAC CTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACC AGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAA GAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAA GACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGA AGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGT TTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAG CCCTGCCACCTAGGTAA (CAR2.3)CAGGTGCAGTTGCAGGAAAGCGGGCCTGG 199 QVQLQESGPGLVKPSQ 200 Clone 24C8CCTTGTGAAACCAAGCCAGACACTGAGCC TLSLTCTVSGGSISSG CD8 CARTGACATGCACTGTGTCCGGCGGGTCCATA GFYWSWIRQHPGKGLE DNA HxLTCTTCCGGGGGTTTTTATTGGTCCTGGAT WIGYIHHSGSTHYNPSACGCCAGCATCCCGGGAAAGGACTTGAAT LKSRVTISIDTSKNLFGGATTGGATATATCCACCATTCCGGAAGC SLRLSSVTAADTAVYYACCCACTACAATCCAAGCCTTAAATCCCG CASLVYCGGDCYSGFDGGTGACAATCTCCATCGACACCTCAAAGA YWGQGTLVTVSSGGGGATCTTTTTTCCCTGCGGTTGTCTTCAGTA SGGGGSGGGGSDIQLTACTGCCGCCGATACCGCTGTGTACTACTG QSPSSLSASVGDRVSFTGCCAGCCTCGTCTATTGCGGCGGAGATT TCQASQDINNFLNWYQGTTATTCTGGGTTCGATTATTGGGGTCAA QKPGKAPKLLIYDASNGGCACACTGGTAACTGTCAGCAGCGGAGG LETGVPSRFSGSGSGTCGGCGGTTCCGGGGGCGGGGGCAGTGGAG DFTFTISSLQPEDIATGGGGCGGATCTGACATTCAGCTTACGCAG YYCQQYGNLPFTFGGGTCCCCATCTTCACTTAGCGCCAGCGTTGG TKVEIKRAAALSNSIMCGATCGGGTCAGCTTCACGTGTCAAGCAA YFSHFVPVFLPAKPTTGTCAGGATATCAACAACTTTCTTAACTGG TPAPRPPTPAPTIASQTACCAGCAGAAGCCAGGCAAGGCACCCAA PLSLRPEACRPAAGGAGTTGCTGATTTACGATGCTTCTAACCTCG VHTRGLDFACDIYIWAAGACGGGAGTGCCTAGCCGCTTCTCCGGG PLAGTCGVLLLSLVITAGCGGCAGCGGCACAGACTTTACCTTTAC LYCNHRNRSKRSRLLHGATTTCCAGTCTGCAGCCAGAGGATATAG SDYMNMTPRRPGPTRKCAACTTATTACTGTCAGCAGTATGGCAAC HYQPYAPPRDFAAYRSCTCCCTTTTACCTTCGGTGGTGGCACAAA RVKFSRSADAPAYQQGGGTCGAGATTAAAAGAGCCGCAGCGTTGT QNQLYNELNLGRREEYCCAACTCCATAATGTATTTTTCTCATTTT DVLDKRRGRDPEMGGKGTGCCCGTCTTTCTGCCTGCCAAACCTAC PRRKNPQEGLYNELQKCACCACCCCCGCCCCACGACCACCTACTC DKMAEAYSEIGMKGERCAGCCCCCACCATCGCCTCCCAGCCCCTC RRGKGHDGLYQGLSTAAGCCTGAGGCCAGAGGCTTGTCGCCCTGC TKDTYDALHMQALPPRTGCGGGGGGCGCTGTCCATACCAGAGGAC TCGACTTCGCCTGCGATATTTATATATGGGCCCCCCTCGCCGGCACCTGCGGAGTCTT GCTCCTGAGCCTTGTGATCACGCTTTATTGTAACCATCGGAATAGATCCAAAAGAAGC CGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGA AACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAA GTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAAC GAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGG ACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGA GCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGG AGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATA CTTATGACGCTCTCCACATGCAAGCCCTG CCACCTAGG(CAR3.1) ATGGCACTCCCCGTAACTGCTCTGCTGCT 201 MALPVTALLLPLALLL 202 CloneGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVQSGAEV 20C5.1 THDGCCCGCAGGTCCAACTGGTGCAGTCCGGA KKPGASVKVSCKVSGY CAR DNAGCCGAAGTCAAGAAACCAGGTGCCTCCGT TLTELSMHWVRQAPGK HxLTAAAGTGAGTTGCAAAGTCTCTGGATACA GLEWMGGFDPEDGETICTCTGACCGAGCTCTCTATGCACTGGGTC YAQKFQGRVTVTEDTSCGGCAGGCCCCCGGCAAGGGATTGGAATG TDTAYMELSSLRSEDTGATGGGCGGGTTCGATCCTGAGGACGGAG AVYYCATESRGIGWPYAGACTATCTACGCTCAAAAATTCCAGGGA FDYWGQGTLVTVSSGGCGAGTGACTGTGACCGAAGACACTAGTAC GGSGGGGSGGGGSDIQCGACACTGCCTACATGGAACTTTCCTCTC MTQSPSSLSASVGDRVTGCGATCAGAAGATACCGCAGTGTACTAC TITCRASQSISSYLNWTGTGCTACTGAATCTAGGGGCATTGGATG YQQKPGKAPKLLISGAGCCCTACTTCGATTACTGGGGTCAGGGAA SSLKSGVPSRFSGSGSCTCTGGTGACTGTCTCCAGCGGTGGAGGT GTDFTLTISSLPPEDFGGCAGCGGTGGTGGCGGAAGCGGGGGGGG ATYYCQQSYSTPITFGCGGCTCTGATATTCAGATGACTCAATCTC QGTRLEIKRAAALDNECTTCTTCTCTGTCCGCTTCCGTGGGCGAT KSNGTIIHVKGKHLCPAGAGTGACCATTACTTGTAGGGCGTCCCA SPLFPGPSKPFWVLVVGTCAATCTCCAGTTATTTGAATTGGTATC VGGVLACYSLLVTVAFAGCAGAAGCCCGGGAAAGCACCTAAGCTG IIFWVRSKRSRLLHSDTTGATCAGCGGGGCTTCTAGCCTGAAGAG YMNMTPRRPGPTRKHYTGGGGTACCTTCACGGTTCAGCGGAAGCG QPYAPPRDFAAYRSRVGAAGCGGAACCGATTTCACCCTGACTATC KFSRSADAPAYQQGQNAGCAGCCTGCCACCTGAGGACTTTGCAAC QLYNELNLGRREEYDVTTACTACTGCCAACAGTCATACAGCACTC LDKRRGRDPEMGGKPRCGATCACTTTCGGCCAGGGCACCCGGCTC RKNPQEGLYNELQKDKGAAATCAAGCGCGCTGCTGCTTTGGACAA MAEAYSEIGMKGERRRTGAGAAGTCAAACGGCACCATCATACATG GKGHDGLYQGLSTATKTTAAAGGTAAACATCTGTGTCCCTCCCCG DTYDALHMQALPPRCTGTTCCCCGGCCCTTCCAAACCGTTCTG GGTTCTGGTGGTGGTCGGAGGCGTACTCGCTTGCTATAGTCTGCTGGTAACTGTCGCC TTCATCATCTTTTGGGTGAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGA ATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACC TAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCA GCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAG AGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAG ACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAA GCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTT TGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGC CCTGCCACCTAGGTAA (CAR3.1)CAGGTCCAACTGGTGCAGTCCGGAGCCGA 203 QVQLVQSGAEVKKPGA 204 CloneAGTCAAGAAACCAGGTGCCTCCGTTAAAG SVKVSCKVSGYTLTEL 20C5.1 THDTGAGTTGCAAAGTCTCTGGATACACTCTG SMHWVRQAPGKGLEWM CAR DNAACCGAGCTCTCTATGCACTGGGTCCGGCA GGFDPEDGETIYAQKF HxLGGCCCCCGGCAAGGGATTGGAATGGATGG QGRVTVTEDTSTDTAYGCGGGTTCGATCCTGAGGACGGAGAGACT MELSSLRSEDTAVYYCATCTACGCTCAAAAATTCCAGGGACGAGT ATESRGIGWPYFDYWGGACTGTGACCGAAGACACTAGTACCGACA QGTLVTVSSGGGGSGGCTGCCTACATGGAACTTTCCTCTCTGCGA GGSGGGGSDIQMTQSPTCAGAAGATACCGCAGTGTACTACTGTGC SSLSASVGDRVTITCRTACTGAATCTAGGGGCATTGGATGGCCCT ASQSISSYLNWYQQKPACTTCGATTACTGGGGTCAGGGAACTCTG GKAPKLLISGASSLKSGTGACTGTCTCCAGCGGTGGAGGTGGCAG GVPSRFSGSGSGTDFTCGGTGGTGGCGGAAGCGGGGGGGGCGGCT LTISSLPPEDFATYYCCTGATATTCAGATGACTCAATCTCCTTCT QQSYSTPITFGQGTRLTCTCTGTCCGCTTCCGTGGGCGATAGAGT EIKRAAALDNEKSNGTGACCATTACTTGTAGGGCGTCCCAGTCAA IIHVKGKHLCPSPLFPTCTCCAGTTATTTGAATTGGTATCAGCAG GPSKPFWVLVVVGGVLAAGCCCGGGAAAGCACCTAAGCTGTTGAT ACYSLLVTVAFIIFWVCAGCGGGGCTTCTAGCCTGAAGAGTGGGG RSKRSRLLHSDYMNMTTACCTTCACGGTTCAGCGGAAGCGGAAGC PRRPGPTRKHYQPYAPGGAACCGATTTCACCCTGACTATCAGCAG PRDFAAYRSRVKFSRSCCTGCCACCTGAGGACTTTGCAACTTACT ADAPAYQQGQNQLYNEACTGCCAACAGTCATACAGCACTCCGATC LNLGRREEYDVLDKRRACTTTCGGCCAGGGCACCCGGCTCGAAAT GRDPEMGGKPRRKNPQCAAGCGCGCTGCTGCTTTGGACAATGAGA EGLYNELQKDKMAEAYAGTCAAACGGCACCATCATACATGTTAAA SEIGMKGERRRGKGHDGGTAAACATCTGTGTCCCTCCCCGCTGTT GLYQGLSTATKDTYDACCCCGGCCCTTCCAAACCGTTCTGGGTTC LHMQALPPR TGGTGGTGGTCGGAGGCGTACTCGCTTGCTATAGTCTGCTGGTAACTGTCGCCTTCAT CATCTTTTGGGTGAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATG ACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAG ATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTA TCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTAT GACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAA AAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTA TTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTAC CAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGC CACCTAGG (CAR3.2)ATGGCACTCCCCGTAACTGCTCTGCTGCT 205 MALPVTALLLPLALLL 206 CloneGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVQSGAEV 20C5.1 CHDGCCCGCAGGTGCAGCTTGTGCAGAGCGGG KKPGASVKVSCKVSGY CAR DNAGCCGAGGTGAAGAAGCCCGGGGCCAGCGT TLTELSMHWVRQAPGK HxLCAAAGTGTCCTGTAAGGTCAGCGGTTACA GLEWMGGFDPEDGETICCCTCACCGAGCTGAGCATGCACTGGGTA YAQKFQGRVTVTEDTSCGGCAGGCTCCCGGCAAAGGTCTTGAGTG TDTAYMELSSLRSEDTGATGGGTGGATTTGATCCAGAAGATGGAG AVYYCATESRGIGWPYAGACTATCTACGCCCAGAAGTTCCAGGGC FDYWGQGTLVTVSSGGCGGGTCACCGTAACAGAAGACACCTCAAC GGSGGGGSGGGGSDIQTGACACCGCTTACATGGAGCTGAGTTCAC MTQSPSSLSASVGDRVTGCGGTCCGAGGACACGGCCGTGTATTAT TITCRASQSISSYLNWTGTGCCACCGAGAGCCGCGGAATCGGATG YQQKPGKAPKLLISGAGCCTTACTTCGACTACTGGGGACAGGGTA SSLKSGVPSRFSGSGSCACTTGTTACAGTATCATCCGGGGGTGGC GTDFTLTISSLPPEDFGGCTCTGGTGGGGGCGGCTCCGGAGGGGG ATYYCQQSYSTPITFGTGGATCAGATATCCAAATGACTCAAAGTC QGTRLEIKRAAAIEVMCAAGTTCCCTGTCTGCCTCAGTCGGAGAT YPPPYLDNEKSNGTIIAGAGTCACCATAACCTGCAGGGCAAGTCA HVKGKHLCPSPLFPGPGTCCATCTCCTCCTATCTGAACTGGTACC SKPFWVLVVVGGVLACAACAGAAACCTGGAAAGGCGCCTAAGCTC YSLLVTVAFIIFWVRSCTGATCTCCGGAGCCTCATCTTTGAAATC KRSRLLHSDYMNMTPRCGGTGTCCCATCTCGCTTCAGTGGCTCTG RPGPTRKHYQPYAPPRGAAGCGGTACAGATTTTACTTTGACCATT DFAAYRSRVKFSRSADAGCAGCCTCCCACCGGAAGACTTTGCTAC APAYQQGQNQLYNELNATATTACTGCCAGCAGTCTTACTCAACCC LGRREEYDVLDKRRGRCAATCACCTTCGGGCAAGGCACCAGACTC DPEMGGKPRRKNPQEGGAAATAAAAAGAGCAGCTGCTATCGAGGT LYNELQKDKMAEAYSETATGTACCCACCGCCGTACTTGGATAACG IGMKGERRRGKGHDGLAAAAAAGCAATGGGACCATCATTCATGTG YQGLSTATKDTYDALHAAGGGTAAGCACCTTTGCCCTAGCCCACT MQALPPR GTTTCCTGGCCCGAGTAAACCCTTTTGGGTACTTGTGGTCGTCGGCGGCGTGCTGGCC TGCTACTCACTCCTGGTTACCGTCGCATTCATCATCTTTTGGGTGAGATCCAAAAGAA GCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAG GAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTG AAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATA ACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACG GGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAAT GAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGC GGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGA TACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR3.2) CAGGTGCAGCTTGTGCAGAGCGGGGCCGA 207QVQLVQSGAEVKKPGA 208 Clone GGTGAAGAAGCCCGGGGCCAGCGTCAAAGSVKVSCKVSGYTLTEL 20C5.1 CHD TGTCCTGTAAGGTCAGCGGTTACACCCTCSMHWVRQAPGKGLEWM CAR DNA ACCGAGCTGAGCATGCACTGGGTACGGCA GGFDPEDGETIYAQKFHxL GGCTCCCGGCAAAGGTCTTGAGTGGATGG QGRVTVTEDTSTDTAYGTGGATTTGATCCAGAAGATGGAGAGACT MELSSLRSEDTAVYYCATCTACGCCCAGAAGTTCCAGGGCCGGGT ATESRGIGWPYFDYWGCACCGTAACAGAAGACACCTCAACTGACA QGTLVTVSSGGGGSGGCCGCTTACATGGAGCTGAGTTCACTGCGG GGSGGGGSDIQMTQSPTCCGAGGACACGGCCGTGTATTATTGTGC SSLSASVGDRVTITCRCACCGAGAGCCGCGGAATCGGATGGCCTT ASQSIISYLNWYQQKPACTTCGACTACTGGGGACAGGGTACACTT GKAPKLLISGASSLKSGTTACAGTATCATCCGGGGGTGGCGGCTC GVPSRFSGSGSGTDFTTGGTGGGGGCGGCTCCGGAGGGGGTGGAT LTISSLPPEDFATYYCCAGATATCCAAATGACTCAAAGTCCAAGT QQSYSTPITFGQGTRLTCCCTGTCTGCCTCAGTCGGAGATAGAGT EIKRAAAIEVMYPPPYCACCATAACCTGCAGGGCAAGTCAGTCCA LDNEKSNGTIIHVKGKTCTCCTCCTATCTGAACTGGTACCAACAG HLCPSPLFPGPSKPFWAAACCTGGAAAGGCGCCTAAGCTCCTGAT VLVVVGGVLACYSLLVCTCCGGAGCCTCATCTTTGAAATCCGGTG TVAFIIFWVRSKRSRLTCCCATCTCGCTTCAGTGGCTCTGGAAGC LHSDYMNMTPRRPGPTGGTACAGATTTTACTTTGACCATTAGCAG RKHYQPYAPPRDFAAYCCTCCCACCGGAAGACTTTGCTACATATT RSRVKFSRSADAPAYQACTGCCAGCAGTCTTACTCAACCCCAATC QGQNQLYNELNLGRREACCTTCGGGCAAGGCACCAGACTCGAAAT EYDVLDKRRGRDPEMGAAAAAGAGCAGCTGCTATCGAGGTTATGT GKPRRKNPQEGLYNELACCCACCGCCGTACTTGGATAACGAAAAA QKDKMAEAYSEIGMKGAGCAATGGGACCATCATTCATGTGAAGGG ERRRGKGHDGLYQGLSTAAGCACCTTTGCCCTAGCCCACTGTTTC TATKDTYDALHMQALPCTGGCCCGAGTAAACCCTTTTGGGTACTT PR GTGGTCGTCGGCGGCGTGCTGGCCTGCTACTCACTCCTGGTTACCGTCGCATTCATCA TCTTTTGGGTGAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGAC TCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGAT TTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATC AGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGA CGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAA AACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATT CTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCA GGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCA CCTAGG (CAR3.3)ATGGCACTCCCCGTAACTGCTCTGCTGCT 209 MALPVTALLLPLALLL 210 CloneGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVQSGAEV 20C5.1 CD8GCCCGCAGGTGCAGTTGGTGCAAAGCGGC KKPGASVKVSCKVSGY CAR DNAGCAGAAGTTAAGAAACCTGGGGCGTCAGT TLTELSMHWVRQAPGK HxLTAAGGTGTCTTGCAAAGTATCTGGCTATA GLEWMGGFDPEDGETICCCTCACTGAGCTGTCCATGCATTGGGTA YAQKFQGRVTVTEDTSAGGCAGGCTCCTGGAAAGGGGCTCGAATG TDTAYMELSSLRSEDTGATGGGAGGATTTGACCCTGAAGACGGAG AVYYCATESRGIGWPYAGACCATCTACGCCCAGAAATTCCAGGGT FDYWGQGTLVTVSSGGAGAGTAACAGTGACTGAGGACACTAGCAC GGSGGGGSGGGGSDIQTGACACAGCGTACATGGAGCTGAGTTCTC MTQSPSSLSASVGDRVTGAGAAGTGAGGACACAGCCGTTTACTAC TITCRASQSISSYLNWTGCGCTACCGAGTCCAGAGGTATTGGCTG YQQKPGKAPKLLISGAGCCATACTTCGACTATTGGGGTCAGGGCA SSLKSGVPSRFSGSGSCCCTGGTTACAGTGAGTTCAGGAGGCGGG GTDFTLTISSLPPEDFGGCTCTGGGGGGGGCGGTTCCGGAGGGGG ATYYCQQSYSTPITFGGGGCTCAGATATACAGATGACGCAGAGTC QGTRLEIKRAAALSNSCATCAAGTCTCTCAGCCAGCGTGGGAGAT IMYFSHFVPVFLPAKPCGCGTGACTATTACTTGCCGCGCCAGCCA TTTPAPRPPTPAPTIAGAGTATTAGCTCCTATCTGAATTGGTACC SQPLSLRPEACRPAAGAGCAAAAGCCCGGGAAGGCCCCTAAGCTT GAVHTRGLDFACDIYICTGATTTCTGGCGCCTCCTCTTTGAAGTC WAPLAGTCGVLLLSLVAGGTGTGCCAAGCAGATTTAGCGGGTCTG ITLYCNHRNRSKRSRLGAAGTGGCACTGACTTTACACTTACTATC LHSDYMNMTPRRPGPTTCCAGCCTGCCCCCAGAGGATTTTGCCAC RKHYQPYAPPRDFAAYATATTACTGTCAGCAAAGCTACTCTACTC RSRVKFSRSADAPAYQCAATCACTTTCGGCCAGGGCACAAGATTG QGQNQLYNELNLGRREGAGATTAAGAGGGCTGCCGCACTTTCAAA EYDVLDKRRGRDPEMGTTCCATCATGTATTTCAGCCATTTTGTGC GKPRRKNPQEGLYNELCTGTTTTTCTTCCGGCCAAACCTACAACC QKDKMAEAYSEIGMKGACTCCCGCCCCACGCCCACCTACTCCCGC ERRRGKGHDGLYQGLSCCCTACCATTGCCTCCCAGCCTCTGTCTC TATKDTYDALHMQALPTTAGACCTGAGGCTTGTAGACCTGCTGCC PR GGCGGAGCCGTGCACACTCGCGGTCTGGACTTCGCCTGCGACATCTATATCTGGGCCC CTCTGGCCGGCACCTGCGGCGTTCTCCTTCTCTCACTCGTAATCACACTCTATTGCAA TCACAGGAACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACT CCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATT TCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCA GCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGAC GTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAA ACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTC TGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAG GGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCAC CTAGGTAA (CAR3.3)CAGGTGCAGTTGGTGCAAAGCGGCGCAGA 211 QVQLVQSGAEVKKPGA 212 CloneAGTTAAGAAACCTGGGGCGTCAGTTAAGG SVKVSCKVSGYTLTEL 20C5.1 CD8TGTCTTGCAAAGTATCTGGCTATACCCTC SMHWVRQAPGKGLEWM CAR DNAACTGAGCTGTCCATGCATTGGGTAAGGCA GGFDPEDGETIYAQKF HxLGGCTCCTGGAAAGGGGCTCGAATGGATGG QGRVTVTEDTSTDTAYGAGGATTTGACCCTGAAGACGGAGAGACC MELSSLRSEDTAVYYCATCTACGCCCAGAAATTCCAGGGTAGAGT ATESRGIGWPYFDYWGAACAGTGACTGAGGACACTAGCACTGACA QGTLVTVSSGGGGSGGCAGCGTACATGGAGCTGAGTTCTCTGAGA GGSGGGGSDIQMTQSPAGTGAGGACACAGCCGTTTACTACTGCGC SSLSASVGDRVTITCRTACCGAGTCCAGAGGTATTGGCTGGCCAT ASQSIISYLNWYQQKPACTTCGACTATTGGGGTCAGGGCACCCTG GKAPKLLISGASSLKSGTTACAGTGAGTTCAGGAGGCGGGGGCTC GVPSRFSGSGSGTDFTTGGGGGGGGCGGTTCCGGAGGGGGGGGCT LTISSLPPEDFATYYCCAGATATACAGATGACGCAGAGTCCATCA QQSYSTPITFGQGTRLAGTCTCTCAGCCAGCGTGGGAGATCGCGT EIKRAAALSNSIMYFSGACTATTACTTGCCGCGCCAGCCAGAGTA HFVPVFLPAKPTTTPATTAGCTCCTATCTGAATTGGTACCAGCAA PRPPTPAPTIASQPLSAAGCCCGGGAAGGCCCCTAAGCTTCTGAT LRPEACRPAAGGAVHTTTCTGGCGCCTCCTCTTTGAAGTCAGGTG RGLDFACDIYIWAPLATGCCAAGCAGATTTAGCGGGTCTGGAAGT GTCGVLLLSLVITLYCGGCACTGACTTTACACTTACTATCTCCAG NHRNRSKRSRLLHSDYCCTGCCCCCAGAGGATTTTGCCACATATT MNMTPRRPGPTRKHYQACTGTCAGCAAAGCTACTCTACTCCAATC PYAPPRDFAAYRSRVKACTTTCGGCCAGGGCACAAGATTGGAGAT FSRSADAPAYQQGQNQTAAGAGGGCTGCCGCACTTTCAAATTCCA LYNELNLGRREEYDVLTCATGTATTTCAGCCATTTTGTGCCTGTT DKRRGRDPEMGGKPRRTTTCTTCCGGCCAAACCTACAACCACTCC KNPQEGLYNELQKDKMCGCCCCACGCCCACCTACTCCCGCCCCTA AEAYSEIGMKGERRRGCCATTGCCTCCCAGCCTCTGTCTCTTAGA KGHDGLYQGLSTATKDCCTGAGGCTTGTAGACCTGCTGCCGGCGG TYDALHMQALPPRAGCCGTGCACACTCGCGGTCTGGACTTCG CCTGCGACATCTATATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTTCTCCTTCTCTC ACTCGTAATCACACTCTATTGCAATCACAGGAACAGATCCAAAAGAAGCCGCCTGCTC CATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACC AGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAG ATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAAC CTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGA TGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAA GGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGA AAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACG CTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.1)ATGGCACTCCCCGTAACTGCTCTGCTGCT 213 MALPVTALLLPLALLL 214 CloneGCCGTTGGCATTGCTCCTGCACGCCGCAC HAARPQVQLVESGGGV 20C5.2 THDGCCCGCAGGTCCAGTTGGTCGAAAGTGGC VQPGRSLRLSCAASGF CAR DNAGGTGGTGTAGTGCAGCCGGGCCGCAGTTT TFSSYGMHWVRQAPGK HxLGAGGCTTTCCTGTGCGGCTTCAGGCTTTA GLEWVAVISYDGSDKYCTTTTTCCAGCTATGGAATGCACTGGGTG YVDSVKGRFTISRDNSCGGCAGGCCCCCGGCAAAGGACTTGAGTG KNRLYLQMNSLRAEDTGGTGGCCGTCATTTCTTATGACGGATCAG AVYYCARERYSGRDYWATAAGTACTACGTGGACAGCGTCAAGGGC GQGTLVTVSSGGGGSGAGATTCACCATCTCTAGGGACAACAGTAA GGGSGGGGSEIVMTQSAAATAGACTCTACCTCCAGATGAATAGCC PATLSVSPGERATLSCTCAGAGCTGAAGACACGGCCGTCTACTAT RASQSVSSLLTWYQQKTGTGCTCGGGAGCGGTATAGTGGCAGAGA PGQAPRLLIFGASTRACTACTGGGGGCAGGGCACACTCGTTACAG TGIPARFSGSGSGTGFTGAGTAGCGGCGGAGGAGGGAGTGGGGGC TLTISSLQSEDFAVYYGGTGGCTCCGGTGGAGGAGGTTCTGAGAT CQQYDTWPFTFGPGTKTGTTATGACCCAGAGTCCTGCGACCCTCT VDFKRAAALDNEKSNGCAGTCAGCCCCGGGGAGCGCGCAACTTTG TIIHVKGKHLCPSPLFTCTTGCAGAGCTAGTCAGTCCGTGTCCTC PGPSKPFWVLVVVGGVTCTTCTGACATGGTACCAGCAAAAGCCCG LACYSLLVTVAFIIFWGGCAGGCTCCGCGCCTTTTGATCTTTGGG VRSKRSRLLHSDYMNMGCTTCAACAAGAGCCACTGGGATTCCCGC TPRRPGPTRKHYQPYAACGATTCTCTGGCTCCGGGAGCGGTACTG PPRDFAAYRSRVKFSRGTTTCACCCTGACGATTAGCAGTCTCCAG SADAPAYQQGQNQLYNAGCGAGGACTTCGCCGTATACTACTGCCA ELNLGRREEYDVLDKRGCAGTACGATACGTGGCCATTCACTTTTG RGRDPEMGGKPRRKNPGACCAGGGACTAAAGTGGATTTTAAGCGC QEGLYNELQKDKMAEAGCCGCCGCTCTCGATAACGAAAAGTCAAA YSEIGMKGERRRGKGHTGGCACCATAATCCACGTCAAAGGCAAGC DGLYQGLSTATKDTYDACCTGTGCCCTTCCCCGCTCTTCCCCGGA ALHMQALPPR CCCAGTAAACCATTTTGGGTGCTGGTTGTTGTGGGGGGCGTGCTGGCCTGCTATAGCC TTTTGGTCACTGTAGCCTTCATTATTTTTTGGGTCAGATCCAAAAGAAGCCGCCTGCT CCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTAC CAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCA GATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAA CCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAG ATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGA AGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGG AAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGAC GCTCTCCACATGCAAGCCCTGCCACCTAG GTAA(CAR4.1) CAGGTCCAGTTGGTCGAAAGTGGCGGTGG 215 QVQLVESGGGVVQPGR 216 CloneTGTAGTGCAGCCGGGCCGCAGTTTGAGGC SLRLSCAASGFTFSSY 20C5.2 THDTTTCCTGTGCGGCTTCAGGCTTTACTTTT GMHWVRQAPGKGLEWV CAR DNATCCAGCTATGGAATGCACTGGGTGCGGCA AVISYDGSDKYYVDSV HxLGGCCCCCGGCAAAGGACTTGAGTGGGTGG KGRFTISRDNSKNRLYCCGTCATTTCTTATGACGGATCAGATAAG LQMNSLRAEDTAVYYCTACTACGTGGACAGCGTCAAGGGCAGATT ARERYSGRDYWGQGTLCACCATCTCTAGGGACAACAGTAAAAATA VTVSSGGGGSGGGGSGGACTCTACCTCCAGATGAATAGCCTCAGA GGGSEIVMTQSPATLSGCTGAAGACACGGCCGTCTACTATTGTGC VSPGERATLSCRASQSTCGGGAGCGGTATAGTGGCAGAGACTACT VSSLLTWYQQKPGQAPGGGGGCAGGGCACACTCGTTACAGTGAGT RLLIFGASTRATGIPAAGCGGCGGAGGAGGGAGTGGGGGCGGTGG RFSGSGSGTGFTLTISCTCCGGTGGAGGAGGTTCTGAGATTGTTA SLQSEDFAVYYCQQYDTGACCCAGAGTCCTGCGACCCTCTCAGTC TWPFTFGPGTKVDFKRAGCCCCGGGGAGCGCGCAACTTTGTCTTG AAALDNEKSNGTIIHVCAGAGCTAGTCAGTCCGTGTCCTCTCTTC KGKHLCPSPLFPGPSKTGACATGGTACCAGCAAAAGCCCGGGCAG PFWVLVVVGGVLACYSGCTCCGCGCCTTTTGATCTTTGGGGCTTC LLVTVAFIIFWVRSKRAACAAGAGCCACTGGGATTCCCGCACGAT SRLLHSDYMNMTPRRPTCTCTGGCTCCGGGAGCGGTACTGGTTTC GPTRKHYQPYAPPRDFACCCTGACGATTAGCAGTCTCCAGAGCGA AAYRSRVKFSRSADAPGGACTTCGCCGTATACTACTGCCAGCAGT AYQQGQNQLYNELNLGACGATACGTGGCCATTCACTTTTGGACCA RREEYDVLDKRRGRDPGGGACTAAAGTGGATTTTAAGCGCGCCGC EMGGKPRRKNPQEGLYCGCTCTCGATAACGAAAAGTCAAATGGCA NELQKDKMAEAYSEIGCCATAATCCACGTCAAAGGCAAGCACCTG MKGERRRGKGHDGLYQTGCCCTTCCCCGCTCTTCCCCGGACCCAG GLSTATKDTYDALHMQTAAACCATTTTGGGTGCTGGTTGTTGTGG ALPPR GGGGCGTGCTGGCCTGCTATAGCCTTTTGGTCACTGTAGCCTTCATTATTTTTTGGGT CAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGC CCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCT ATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCA GAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGAC AAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGG AGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGG CATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGC ACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.2) ATGGCACTCCCCGTAACTGCTCTGCTGCT 217MALPVTALLLPLALLL 218 Clone GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPQVQLVESGGGV 20C5.2 CHD GCCCGCAGGTGCAGCTCGTGGAGTCTGGCVQPGRSLRLSCAASGF CAR DNA GGCGGCGTGGTCCAGCCCGGCCGGTCCCT TFSSYGMHWVRQAPGKHxL GCGCCTGTCCTGCGCCGCCAGCGGGTTTA GLEWVAVISYDGSDKYCTTTTTCCTCCTACGGCATGCACTGGGTG YVDSVKGRFTISRDNSCGCCAGGCTCCCGGCAAGGGCCTCGAGTG KNRLYLQMNSLRAEDTGGTCGCCGTGATCTCATACGATGGGTCAG AVYYCARERYSGRDYWACAAATACTATGTCGATTCTGTTAAAGGG GQGTLVTVSSGGGGSGCGGTTTACCATTTCAAGAGATAACTCTAA GGGSGGGGSEIVMTQSGAATAGGCTGTATTTGCAGATGAACAGCC PATLSVSPGERATLSCTGAGGGCTGAAGATACCGCAGTGTACTAT RASQSVSSLLTWYQQKTGCGCTAGGGAGCGGTATAGTGGCCGCGA PGQAPRLLIFGASTRATTACTGGGGACAGGGTACACTGGTGACCG TGIPARFSGSGSGTGFTGAGCTCTGGGGGTGGCGGAAGCGGGGGT TLTISSLQSEDFAVYYGGCGGAAGCGGCGGAGGGGGTAGTGAAAT CQQYDTWPFTFGPGTKTGTGATGACCCAGTCTCCGGCTACACTTT VDFKRAAAIEVMYPPPCAGTCTCCCCTGGGGAGAGAGCTACACTG YLDNEKSNGTIIHVKGTCATGCAGAGCGTCCCAGTCCGTCTCTTC KHLCPSPLFPGPSKPFTCTCCTTACCTGGTATCAGCAGAAGCCCG WVLVVVGGVLACYSLLGCCAGGCTCCTCGACTGCTGATCTTCGGT VTVAFIIFWVRSKRSRGCCTCCACAAGGGCGACCGGGATTCCAGC LLHSDYMNMTPRRPGPCCGCTTCTCAGGTTCTGGGAGCGGAACTG TRKHYQPYAPPRDFAAGTTTCACTTTGACAATCAGTTCACTGCAG YRSRVKFSRSADAPAYTCAGAGGATTTCGCCGTGTACTACTGCCA QQGQNQLYNELNLGRRGCAATACGACACATGGCCATTCACTTTCG EEYDVLDKRRGRDPEMGACCCGGTACCAAAGTCGATTTCAAGAGA GGKPRRKNPQEGLYNEGCCGCGGCCATCGAGGTTATGTACCCACC LQKDKMAEAYSEIGMKACCATATCTGGACAATGAAAAAAGCAATG GERRRGKGHDGLYQGLGAACCATTATCCATGTGAAGGGTAAACAC STATKDTYDALHMQALCTCTGCCCTAGCCCACTTTTCCCTGGCCC PPR ATCAAAGCCCTTCTGGGTCTTGGTGGTCGTGGGGGGTGTGCTGGCCTGTTACAGCCTT CTGGTGACGGTTGCTTTCATTATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCC ATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCA GCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGA TCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACC TGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGAT GGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAG GATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAA AAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGC TCTCCACATGCAAGCCCTGCCACCTAGGT AA (CAR4.2)CAGGTGCAGCTCGTGGAGTCTGGCGGCGG 219 QVQLVESGGGVVQPGR 220 CloneCGTGGTCCAGCCCGGCCGGTCCCTGCGCC SLRLSCAASGFTFSSY 20C5.2 CHDTGTCCTGCGCCGCCAGCGGGTTTACTTTT GMHWVRQAPGKGLEWV CAR DNATCCTCCTACGGCATGCACTGGGTGCGCCA AVISYDGSDKYYVDSV HxLGGCTCCCGGCAAGGGCCTCGAGTGGGTCG KGRFTISRDNSKNRLYCCGTGATCTCATACGATGGGTCAGACAAA LQMNSLRAEDTAVYYCTACTATGTCGATTCTGTTAAAGGGCGGTT ARERYSGRDYWGQGTLTACCATTTCAAGAGATAACTCTAAGAATA VTVSSGGGGSGGGGSGGGCTGTATTTGCAGATGAACAGCCTGAGG GGGSEIVMTQSPATLSGCTGAAGATACCGCAGTGTACTATTGCGC VSPGERATLSCRASQSTAGGGAGCGGTATAGTGGCCGCGATTACT VSSLLTWYQQKPGQAPGGGGACAGGGTACACTGGTGACCGTGAGC RLLIFGASTRATGIPATCTGGGGGTGGCGGAAGCGGGGGTGGCGG RFSGSGSGTGFTLTISAAGCGGCGGAGGGGGTAGTGAAATTGTGA SLQSEDFAVYYCQQYDTGACCCAGTCTCCGGCTACACTTTCAGTC TWPFTFGPGTKVDFKRTCCCCTGGGGAGAGAGCTACACTGTCATG AAAIEVMYPPPYLDNECAGAGCGTCCCAGTCCGTCTCTTCTCTCC KSNGTIIHVKGKHLCPTTACCTGGTATCAGCAGAAGCCCGGCCAG SPLFPGPSKPFWVLVVGCTCCTCGACTGCTGATCTTCGGTGCCTC VGGVLACYSLLVTVAFCACAAGGGCGACCGGGATTCCAGCCCGCT IIFWVRSKRSRLLHSDTCTCAGGTTCTGGGAGCGGAACTGGTTTC YMNMTPRRPGPTRKHYACTTTGACAATCAGTTCACTGCAGTCAGA QPYAPPRDFAAYRSRVGGATTTCGCCGTGTACTACTGCCAGCAAT KFSRSADAPAYQQGQNACGACACATGGCCATTCACTTTCGGACCC QLYNELNLGRREEYDVGGTACCAAAGTCGATTTCAAGAGAGCCGC LDKRRGRDPEMGGKPRGGCCATCGAGGTTATGTACCCACCACCAT RKNPQEGLYNELQKDKATCTGGACAATGAAAAAAGCAATGGAACC MAEAYSEIGMKGERRRATTATCCATGTGAAGGGTAAACACCTCTG GKGHDGLYQGLSTATKCCCTAGCCCACTTTTCCCTGGCCCATCAA DTYDALHMQALPPRAGCCCTTCTGGGTCTTGGTGGTCGTGGGG GGTGTGCTGGCCTGTTACAGCCTTCTGGTGACGGTTGCTTTCATTATCTTCTGGGTTA GATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCC TGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTAT CGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGA ACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAA GCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAG GGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCA TGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCAC TGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.3) ATGGCACTCCCCGTAACTGCTCTGCTGCT 221MALPVTALLLPLALLL 222 Clone GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPQVQLVESGGGV 20C5.2 CD8 GCCCGCAGGTGCAGTTGGTTGAATCAGGAVQPGRSLRLSCAASGF CAR DNA GGGGGTGTGGTGCAACCCGGTCGGTCACT TFSSYGMHWVRQAPGKHxL GCGCCTCAGTTGTGCTGCTTCCGGGTTTA GLEWVAVISYDGSDKYCTTTCAGCTCATATGGGATGCACTGGGTA YVDSVKGRFTISRDNSCGGCAGGCTCCAGGTAAAGGCTTGGAATG KNRLYLQMNSLRAEDTGGTGGCGGTGATCAGCTATGACGGCTCTG AVYYCARERYSGRDYWACAAATATTATGTGGACTCCGTGAAAGGC GQGTLVTVSSGGGGSGAGATTCACCATCAGTCGAGACAACTCAAA GGGSGGGGSEIVMTQSGAATAGACTCTACTTGCAGATGAATAGCC PATLSVSPGERATLSCTCCGGGCCGAAGATACTGCAGTCTATTAT RASQSVSSLLTWYQQKTGCGCCCGGGAGCGCTACAGTGGAAGAGA PGQAPRLLIFGASTRACTATTGGGGGCAAGGAACTCTTGTCACAG TGIPARFSGSGSGTGFTCTCATCTGGCGGCGGCGGCAGCGGTGGG TLTISSLQSEDFAVYYGGCGGATCTGGCGGGGGCGGCAGCGAAAT CQQYDTWPFTFGPGTKCGTTATGACTCAGAGTCCTGCCACACTGA VDFKRAAALSNSIMYFGCGTTAGCCCTGGTGAGAGAGCAACACTT SHFVPVFLPAKPTTTPAGCTGCAGAGCTAGTCAGAGTGTTTCCAG APRPPTPAPTIASQPLTCTTTTGACATGGTACCAACAGAAGCCCG SLRPEACRPAAGGAVHGTCAAGCTCCACGACTGCTCATCTTCGGT TRGLDFACDIYIWAPLGCATCCACCCGCGCAACCGGGATACCCGC AGTCGVLLLSLVITLYCCGGTTTTCCGGTTCTGGAAGTGGCACAG CNHRNRSKRSRLLHSDGATTCACGCTCACCATTTCTTCTCTGCAG YMNMTPRRPGPTRKHYTCTGAAGACTTTGCCGTGTATTACTGCCA QPYAPPRDFAAYRSRVGCAGTACGATACCTGGCCCTTTACCTTTG KFSRSADAPAYQQGQNGCCCAGGTACTAAAGTGGATTTTAAACGA QLYNELNLGRREEYDVGCTGCTGCACTTTCCAATAGTATTATGTA LDKRRGRDPEMGGKPRCTTTTCACATTTTGTGCCCGTGTTCCTGC RKNPQEGLYNELQKDKCTGCGAAGCCTACGACAACCCCAGCCCCT MAEAYSEIGMKGERRRAGGCCGCCCACACCGGCCCCAACTATTGC GKGHDGLYQGLSTATKCTCCCAGCCATTGTCTCTGAGACCCGAAG DTYDALHMQALPPRCTTGCAGACCTGCTGCTGGAGGCGCCGTT CACACCCGAGGATTGGATTTCGCATGTGACATTTACATCTGGGCCCCTTTGGCCGGAA CCTGCGGTGTGCTGCTGCTGTCACTCGTGATTACACTTTACTGCAACCACCGAAACAG ATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCT GGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATC GGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAA CCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAG CGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGG GTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCAT GAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACT GCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR4.3) CAGGTGCAGTTGGTTGAATCAGGAGGGGG 223QVQLVESGGGVVQPGR 224 Clone TGTGGTGCAACCCGGTCGGTCACTGCGCCSLRLSCAASGFTFSSY 20C5.2 CD8 TCAGTTGTGCTGCTTCCGGGTTTACTTTCGMHWVRQAPGKGLEWV CAR DNA AGCTCATATGGGATGCACTGGGTACGGCA AVISYDGSDKYYVDSVHxL GGCTCCAGGTAAAGGCTTGGAATGGGTGG KGRFTISRDNSKNRLYCGGTGATCAGCTATGACGGCTCTGACAAA LQMNSLRAEDTAVYYCTATTATGTGGACTCCGTGAAAGGCAGATT ARERYSGRDYWGQGTLCACCATCAGTCGAGACAACTCAAAGAATA VTVSSGGGGSGGGGSGGACTCTACTTGCAGATGAATAGCCTCCGG GGGSEIVMTQSPATLSGCCGAAGATACTGCAGTCTATTATTGCGC VSPGERATLSCRASQSCCGGGAGCGCTACAGTGGAAGAGACTATT VSSLLTWYQQKPGQAPGGGGGCAAGGAACTCTTGTCACAGTCTCA RLLIFGASTRATGIPATCTGGCGGCGGCGGCAGCGGTGGGGGCGG RFSGSGSGTGFTLTISATCTGGCGGGGGCGGCAGCGAAATCGTTA SLQSEDFAVYYCQQYDTGACTCAGAGTCCTGCCACACTGAGCGTT TWPFTFGPGTKVDFKRAGCCCTGGTGAGAGAGCAACACTTAGCTG AAALSNSIMYFSHFVPCAGAGCTAGTCAGAGTGTTTCCAGTCTTT VFLPAKPTTTPAPRPPTGACATGGTACCAACAGAAGCCCGGTCAA TPAPTIASQPLSLRPEGCTCCACGACTGCTCATCTTCGGTGCATC ACRPAAGGAVHTRGLDCACCCGCGCAACCGGGATACCCGCCCGGT FACDIYIWAPLAGTCGTTTCCGGTTCTGGAAGTGGCACAGGATTC VLLLSLVITLYCNHRNACGCTCACCATTTCTTCTCTGCAGTCTGA RSKRSRLLHSDYMNMTAGACTTTGCCGTGTATTACTGCCAGCAGT PRRPGPTRKHYQPYAPACGATACCTGGCCCTTTACCTTTGGCCCA PRDFAAYRSRVKFSRSGGTACTAAAGTGGATTTTAAACGAGCTGC ADAPAYQQGQNQLYNETGCACTTTCCAATAGTATTATGTACTTTT LNLGRREEYDVLDKRRCACATTTTGTGCCCGTGTTCCTGCCTGCG GRDPEMGGKPRRKNPQAAGCCTACGACAACCCCAGCCCCTAGGCC EGLYNELQKDKMAEAYGCCCACACCGGCCCCAACTATTGCCTCCC SEIGMKGERRRGKGHDAGCCATTGTCTCTGAGACCCGAAGCTTGC GLYQGLSTATKDTYDAAGACCTGCTGCTGGAGGCGCCGTTCACAC LHMQALPPR CCGAGGATTGGATTTCGCATGTGACATTTACATCTGGGCCCCTTTGGCCGGAACCTGC GGTGTGCTGCTGCTGTCACTCGTGATTACACTTTACTGCAACCACCGAAACAGATCCA AAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCC CACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAAC TGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAG AGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTC TATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAG GAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTAC GAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.4) ATGGCACTCCCCGTAACTGCTCTGCTGCT 225MALPVTALLLPLALLL 226 Clone GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPEIVMTQSPATL 20C5.2 THD GCCCGGAGATTGTGATGACCCAGTCCCCTSVSPGERATLSCRASQ CAR DNA GCTACCCTGTCCGTCAGTCCGGGCGAGAG SVSSLLTWYQQKPGQALxH AGCCACCTTGTCATGCCGGGCCAGCCAGT PRLLIFGASTRATGIPCCGTCAGCAGTCTCCTGACTTGGTATCAG ARFSGSGSGTGFTLTICAAAAACCAGGGCAGGCACCGCGGCTTTT SSLQSEDFAVYYCQQYGATTTTTGGTGCAAGCACACGCGCCACTG DTWPFTFGPGTKVDFKGCATTCCAGCTAGGTTTTCTGGAAGTGGA RGGGGSGGGGSGGGGSTCTGGGACAGGCTTCACTCTGACAATCAG QVQLVESGGGVVQPGRTAGCCTGCAGAGTGAGGACTTTGCTGTTT SLRLSCAASGFTFSSYACTACTGTCAACAGTACGACACCTGGCCA GMHWVRQAPGKGLEWVTTCACATTCGGGCCCGGCACCAAGGTCGA AVISYDGSDKYYVDSVCTTCAAGAGGGGCGGTGGAGGTTCAGGTG KGRFTISRDNSKNRLYGTGGCGGGTCAGGCGGCGGTGGGTCTCAG LQMNSLRAEDTAVYYCGTTCAACTGGTGGAATCAGGTGGCGGCGT ARERYSGRDYWGQGTLTGTCCAACCGGGGCGATCACTTCGACTTT VTVSSAAALDNEKSNGCCTGTGCTGCCTCAGGCTTTACTTTTTCA TIIHVKGKHLCPSPLFTCCTATGGGATGCACTGGGTTCGGCAGGC PGPSKPFWVLVVVGGVTCCCGGAAAAGGACTCGAGTGGGTTGCAG LACYSLLVTVAFIIFWTGATCTCTTACGATGGCTCAGACAAGTAT VRSKRSRLLHSDYMNMTATGTGGACTCAGTCAAGGGGAGATTCAC TPRRPGPTRKHYQPYAAATAAGCCGAGACAACTCCAAAAACCGGC PPRDFAAYRSRVKFSRTTTATCTCCAGATGAACAGCCTTAGAGCG SADAPAYQQGQNQLYNGAAGATACCGCGGTATACTACTGTGCCCG ELNLGRREEYDVLDKRCGAGAGGTATTCCGGCAGAGACTACTGGG RGRDPEMGGKPRRKNPGACAGGGCACACTGGTCACCGTGAGTTCT QEGLYNELQKDKMAEAGCCGCAGCGCTCGATAACGAAAAGAGCAA YSEIGMKGERRRGKGHCGGAACCATTATCCACGTTAAGGGCAAGC DGLYQGLSTATKDTYDACCTGTGCCCCAGTCCCCTCTTCCCAGGA ALHMQALPPR CCATCTAAACCCTTCTGGGTTCTGGTAGTAGTTGGAGGGGTCCTTGCATGTTACTCCC TTTTGGTCACCGTCGCCTTCATTATTTTCTGGGTGAGATCCAAAAGAAGCCGCCTGCT CCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTAC CAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCA GATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAA CCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAG ATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGA AGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGG AAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGAC GCTCTCCACATGCAAGCCCTGCCACCTAG GTAA(CAR4.4) GAGATTGTGATGACCCAGTCCCCTGCTAC 227 EIVMTQSPATLSVSPG 228 CloneCCTGTCCGTCAGTCCGGGCGAGAGAGCCA ERATLSCRASQSVSSL 20C5.2 THDCCTTGTCATGCCGGGCCAGCCAGTCCGTC LTWYQQKPGQAPRLLI CAR DNAAGCAGTCTCCTGACTTGGTATCAGCAAAA FGASTRATGIPARFSG LxHACCAGGGCAGGCACCGCGGCTTTTGATTT SGSGTGFTLTISSLQSTTGGTGCAAGCACACGCGCCACTGGCATT EDFAVYYCQQYDTWPFCCAGCTAGGTTTTCTGGAAGTGGATCTGG TFGPGTKVDFKRGGGGGACAGGCTTCACTCTGACAATCAGTAGCC SGGGGSGGGGSQVQLVTGCAGAGTGAGGACTTTGCTGTTTACTAC ESGGGVVQPGRSLRLSTGTCAACAGTACGACACCTGGCCATTCAC CAASGFTFSSYGMHWVATTCGGGCCCGGCACCAAGGTCGACTTCA RQAPGKGLEWVAVISYAGAGGGGCGGTGGAGGTTCAGGTGGTGGC DGSDKYYVDSVKGRFTGGGTCAGGCGGCGGTGGGTCTCAGGTTCA ISRDNSKNRLYLQMNSACTGGTGGAATCAGGTGGCGGCGTTGTCC LRAEDTAVYYCARERYAACCGGGGCGATCACTTCGACTTTCCTGT SGRDYWGQGTLVTVSSGCTGCCTCAGGCTTTACTTTTTCATCCTA AAALDNEKSNGTIIHVTGGGATGCACTGGGTTCGGCAGGCTCCCG KGKHLCPSPLFPGPSKGAAAAGGACTCGAGTGGGTTGCAGTGATC PFWVLVVVGGVLACYSTCTTACGATGGCTCAGACAAGTATTATGT LLVTVAFIIFWVRSKRGGACTCAGTCAAGGGGAGATTCACAATAA SRLLHSDYMNMTPRRPGCCGAGACAACTCCAAAAACCGGCTTTAT GPTRKHYQPYAPPRDFCTCCAGATGAACAGCCTTAGAGCGGAAGA AAYRSRVKFSRSADAPTACCGCGGTATACTACTGTGCCCGCGAGA AYQQGQNQLYNELNLGGGTATTCCGGCAGAGACTACTGGGGACAG RREEYDVLDKRRGRDPGGCACACTGGTCACCGTGAGTTCTGCCGC EMGGKPRRKNPQEGLYAGCGCTCGATAACGAAAAGAGCAACGGAA NELQKDKMAEAYSEIGCCATTATCCACGTTAAGGGCAAGCACCTG MKGERRRGKGHDGLYQTGCCCCAGTCCCCTCTTCCCAGGACCATC GLSTATKDTYDALHMQTAAACCCTTCTGGGTTCTGGTAGTAGTTG ALPPR GAGGGGTCCTTGCATGTTACTCCCTTTTGGTCACCGTCGCCTTCATTATTTTCTGGGT GAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGC CCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCT ATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCA GAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGAC AAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGG AGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGG CATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGC ACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.5) ATGGCACTCCCCGTAACTGCTCTGCTGCT 229MALPVTALLLPLALLL 230 Clone GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPEIVMTQSPATL 20C5.2 CHD GCCCGGAGATCGTCATGACACAGAGTCCASVSPGERATLSCRASQ CAR DNA GCTACCCTGAGCGTGTCCCCTGGAGAGAG SVSSLLTWYQQKPGQALxH AGCCACCCTGTCCTGTAGGGCTAGTCAGA PRLLIFGASTRATGIPGTGTGTCCAGCCTCCTCACCTGGTATCAA ARFSGSGSGTGFTLTICAGAAGCCTGGTCAAGCTCCCCGGCTGCT SSLQSEDFAVYYCQQYTATCTTCGGGGCCAGCACGCGAGCCACAG DTWPFTFGPGTKVDFKGCATCCCGGCCAGATTCTCTGGCTCTGGC RGGGGSGGGGSGGGGSAGTGGCACCGGGTTCACTCTCACGATCTC QVQLVESGGGVVQPGRATCCCTGCAGTCAGAGGATTTCGCTGTGT SLRLSCAASGFTFSSYATTACTGTCAGCAGTACGATACATGGCCC GMHWVRQAPGKGLEWVTTCACCTTCGGCCCGGGCACAAAAGTAGA AVISYDGSDKYYVDSVTTTCAAGCGCGGCGGCGGGGGTAGTGGGG KGRFTISRDNSKNRLYGCGGGGGATCAGGAGGAGGGGGCTCCCAA LQMNSLRAEDTAVYYCGTACAGCTGGTTGAGAGCGGCGGCGGGGT ARERYSGRDYWGQGTLGGTTCAGCCCGGGCGCAGCCTCAGGCTGA VTVSSAAAIEVMYPPPGTTGCGCAGCATCAGGATTCACATTCAGT YLDNEKSNGTIIHVKGTCTTATGGAATGCATTGGGTCAGACAGGC KHLCPSPLFPGPSKPFTCCCGGGAAGGGCCTTGAATGGGTGGCAG WVLVVVGGVLACYSLLTCATTAGCTACGACGGAAGCGATAAGTAC VTVAFIIFWVRSKRSRTATGTGGACTCAGTTAAAGGGAGATTTAC LLHSDYMNMTPRRPGPTATCAGCCGCGACAATTCCAAAAACAGAT TRKHYQPYAPPRDFAATGTATTTGCAGATGAACTCCCTCAGGGCG YRSRVKFSRSADAPAYGAGGACACTGCTGTATATTACTGCGCACG QQGQNQLYNELNLGRRAGAGAGATACTCCGGCCGAGACTATTGGG EEYDVLDKRRGRDPEMGCCAAGGAACATTGGTAACTGTGAGCTCC GGKPRRKNPQEGLYNEGCCGCAGCTATTGAGGTCATGTACCCCCC LQKDKMAEAYSEIGMKACCTTATCTCGATAATGAGAAGAGTAATG GERRRGKGHDGLYQGLGGACTATAATTCACGTAAAGGGCAAACAC STATKDTYDALHMQALCTGTGCCCTTCCCCGCTGTTTCCAGGTCC PPR AAGTAAGCCGTTCTGGGTCCTGGTTGTGGTGGGAGGGGTGCTGGCCTGCTATTCTCTG TTGGTTACCGTGGCCTTTATCATTTTCTGGGTGAGATCCAAAAGAAGCCGCCTGCTCC ATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCA GCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGA TCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACC TGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGAT GGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAG GATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAA AAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGC TCTCCACATGCAAGCCCTGCCACCTAGGT AA (CAR4.5)GAGATCGTCATGACACAGAGTCCAGCTAC 231 EIVMTQSPATLSVSPG 232 CloneCCTGAGCGTGTCCCCTGGAGAGAGAGCCA ERATLSCRASQSVSSL 20C5.2 CHDCCCTGTCCTGTAGGGCTAGTCAGAGTGTG LTWYQQKPGQAPRLLI CAR DNATCCAGCCTCCTCACCTGGTATCAACAGAA FGASTRATGIPARFSG LxHGCCTGGTCAAGCTCCCCGGCTGCTTATCT SGSGTGFTLTISSLQSTCGGGGCCAGCACGCGAGCCACAGGCATC EDFAVYYCQQYDTWPFCCGGCCAGATTCTCTGGCTCTGGCAGTGG TFGPGTKVDFKRGGGGCACCGGGTTCACTCTCACGATCTCATCCC SGGGGSGGGGSQVQLVTGCAGTCAGAGGATTTCGCTGTGTATTAC ESGGGVVQPGRSLRLSTGTCAGCAGTACGATACATGGCCCTTCAC CAASGFTFSSYGMHWVCTTCGGCCCGGGCACAAAAGTAGATTTCA RQAPGKGLEWVAVISYAGCGCGGCGGCGGGGGTAGTGGGGGCGGG DGSDKYYVDSVKGRFTGGATCAGGAGGAGGGGGCTCCCAAGTACA ISRDNSKNRLYLQMNSGCTGGTTGAGAGCGGCGGCGGGGTGGTTC LRAEDTAVYYCARERYAGCCCGGGCGCAGCCTCAGGCTGAGTTGC SGRDYWGQGTLVTVSSGCAGCATCAGGATTCACATTCAGTTCTTA AAAIEVMYPPPYLDNETGGAATGCATTGGGTCAGACAGGCTCCCG KSNGTIIHVKGKHLCPGGAAGGGCCTTGAATGGGTGGCAGTCATT SPLFPGPSKPFWVLVVAGCTACGACGGAAGCGATAAGTACTATGT VGGVLACYSLLVTVAFGGACTCAGTTAAAGGGAGATTTACTATCA IIFWVRSKRSRLLHSDGCCGCGACAATTCCAAAAACAGATTGTAT YMNMTPRRPGPTRKHYTTGCAGATGAACTCCCTCAGGGCGGAGGA QPYAPPRDFAAYRSRVCACTGCTGTATATTACTGCGCACGAGAGA KFSRSADAPAYQQGQNGATACTCCGGCCGAGACTATTGGGGCCAA QLYNELNLGRREEYDVGGAACATTGGTAACTGTGAGCTCCGCCGC LDKRRGRDPEMGGKPRAGCTATTGAGGTCATGTACCCCCCACCTT RKNPQEGLYNELQKDKATCTCGATAATGAGAAGAGTAATGGGACT MAEAYSEIGMKGERRRATAATTCACGTAAAGGGCAAACACCTGTG GKGHDGLYQGLSTATKCCCTTCCCCGCTGTTTCCAGGTCCAAGTA DTYDALHMQALPPRAGCCGTTCTGGGTCCTGGTTGTGGTGGGA GGGGTGCTGGCCTGCTATTCTCTGTTGGTTACCGTGGCCTTTATCATTTTCTGGGTGA GATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCC TGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTAT CGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGA ACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAA GCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAG GGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCA TGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCAC TGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (CAR4.6) ATGGCACTCCCCGTAACTGCTCTGCTGCT 233MALPVTALLLPLALLL 234 Clone GCCGTTGGCATTGCTCCTGCACGCCGCACHAARPEIVMTQSPATL 20C5.2 CD8 GCCCGGAAATAGTGATGACTCAGTCCCCGSVSPGERATLSCRASQ CAR DNA GCCACCCTCAGCGTGTCCCCCGGGGAGCG SVSSLLTWYQQKPGQALxH AGCGACCCTGTCATGCAGGGCTTCCCAGA PRLLIFGASTRATGIPGTGTCAGCTCCCTGCTCACTTGGTATCAG ARFSGSGSGTGFTLTICAAAAGCCGGGGCAGGCTCCCCGCCTCCT SSLQSEDFAVYYCQQYCATCTTCGGGGCATCAACTAGGGCCACCG DTWPFTFGPGTKVDFKGCATTCCTGCAAGATTTTCCGGGTCTGGC RGGGGSGGGGSGGGGSAGCGGCACCGGCTTCACCCTTACCATTAG QVQLVESGGGVVQPGRCTCTCTGCAGTCTGAGGACTTCGCCGTTT SLRLSCAASGFTFSSYACTATTGTCAGCAGTATGATACTTGGCCC GMHWVRQAPGKGLEWVTTTACCTTCGGTCCCGGAACTAAGGTGGA AVISYDGSDKYYVDSVCTTCAAGCGCGGGGGGGGTGGATCTGGAG KGRFTISRDNSKNRLYGTGGTGGCTCCGGGGGCGGTGGAAGCCAG LQMNSLRAEDTAVYYCGTCCAGTTGGTTGAGAGCGGCGGCGGAGT ARERYSGRDYWGQGTLGGTGCAGCCCGGGAGGTCCTTGCGGCTGA VTVSSAAALSNSIMYFGCTGTGCAGCCTCCGGTTTTACTTTTTCT SHFVPVFLPAKPTTTPAGCTATGGAATGCATTGGGTAAGACAGGC APRPPTPAPTIASQPLTCCCGGAAAAGGCCTCGAGTGGGTGGCGG SLRPEACRPAAGGAVHTCATTAGCTATGATGGATCTGATAAATAC TRGLDFACDIYIWAPLTATGTGGACTCAGTTAAGGGGCGCTTCAC AGTCGVLLLSLVITLYAATCTCAAGAGACAATAGCAAAAATAGAC CNHRNRSKRSRLLHSDTGTACCTGCAGATGAATAGTCTGCGCGCC YMNMTPRRPGPTRKHYGAGGACACTGCCGTGTACTACTGCGCCCG QPYAPPRDFAAYRSRVCGAGAGATACAGCGGACGGGATTACTGGG KFSRSADAPAYQQGQNGCCAGGGTACCCTCGTAACGGTGTCCTCC QLYNELNLGRREEYDVGCTGCCGCCCTTAGCAACAGCATTATGTA LDKRRGRDPEMGGKPRCTTTTCTCATTTCGTGCCAGTCTTTCTCC RKNPQEGLYNELQKDKCAGCAAAGCCCACCACTACCCCGGCCCCC MAEAYSEIGMKGERRRAGGCCGCCTACTCCTGCCCCCACTATCGC GKGHDGLYQGLSTATKGTCTCAGCCTCTCTCCTTGCGGCCCGAGG DTYDALHMQALPPRCCTGCCGGCCAGCCGCAGGGGGCGCCGTA CATACTCGGGGTTTGGATTTCGCTTGCGACATATATATTTGGGCCCCCCTCGCCGGCA CATGTGGAGTGCTGCTCCTGAGTCTCGTTATAACCCTCTATTGCAACCATAGAAACAG ATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCT GGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATC GGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAA CCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAG CGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGG GTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCAT GAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACT GCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (CAR4.6) GAAATAGTGATGACTCAGTCCCCGGCCAC 235EIVMTQSPATLSVSPG 236 Clone CCTCAGCGTGTCCCCCGGGGAGCGAGCGAERATLSCRASQSVSSL 20C5.2 CD8 CCCTGTCATGCAGGGCTTCCCAGAGTGTCLTWYQQKPGQAPRLLI CAR DNA AGCTCCCTGCTCACTTGGTATCAGCAAAA FGASTRATGIPARFSGLxH GCCGGGGCAGGCTCCCCGCCTCCTCATCT SGSGTGFTLTISSLQSTCGGGGCATCAACTAGGGCCACCGGCATT EDFAVYYCQQYDTWPFCCTGCAAGATTTTCCGGGTCTGGCAGCGG TFGPGTKVDFKRGGGGCACCGGCTTCACCCTTACCATTAGCTCTC SGGGGSGGGGSQVQLVTGCAGTCTGAGGACTTCGCCGTTTACTAT ESGGGVVQPGRSLRLSTGTCAGCAGTATGATACTTGGCCCTTTAC CAASGFTFSSYGMHWVCTTCGGTCCCGGAACTAAGGTGGACTTCA RQAPGKGLEWVAVISYAGCGCGGGGGGGGTGGATCTGGAGGTGGT DGSDKYYVDSVKGRFTGGCTCCGGGGGCGGTGGAAGCCAGGTCCA ISRDNSKNRLYLQMNSGTTGGTTGAGAGCGGCGGCGGAGTGGTGC LRAEDTAVYYCARERYAGCCCGGGAGGTCCTTGCGGCTGAGCTGT SGRDYWGQGTLVTVSSGCAGCCTCCGGTTTTACTTTTTCTAGCTA AAALSNSIMYFSHFVPTGGAATGCATTGGGTAAGACAGGCTCCCG VFLPAKPTTTPAPRPPGAAAAGGCCTCGAGTGGGTGGCGGTCATT TPAPTIASQPLSLRPEAGCTATGATGGATCTGATAAATACTATGT ACRPAAGGAVHTRGLDGGACTCAGTTAAGGGGCGCTTCACAATCT FACDIYIWAPLAGTCGCAAGAGACAATAGCAAAAATAGACTGTAC VLLLSLVITLYCNHRNCTGCAGATGAATAGTCTGCGCGCCGAGGA RSKRSRLLHSDYMNMTCACTGCCGTGTACTACTGCGCCCGCGAGA PRRPGPTRKHYQPYAPGATACAGCGGACGGGATTACTGGGGCCAG PRDFAAYRSRVKFSRSGGTACCCTCGTAACGGTGTCCTCCGCTGC ADAPAYQQGQNQLYNECGCCCTTAGCAACAGCATTATGTACTTTT LNLGRREEYDVLDKRRCTCATTTCGTGCCAGTCTTTCTCCCAGCA GRDPEMGGKPRRKNPQAAGCCCACCACTACCCCGGCCCCCAGGCC EGLYNELQKDKMAEAYGCCTACTCCTGCCCCCACTATCGCGTCTC SEIGMKGERRRGKGHDAGCCTCTCTCCTTGCGGCCCGAGGCCTGC GLYQGLSTATKDTYDACGGCCAGCCGCAGGGGGCGCCGTACATAC LHMQALPPR TCGGGGTTTGGATTTCGCTTGCGACATATATATTTGGGCCCCCCTCGCCGGCACATGT GGAGTGCTGCTCCTGAGTCTCGTTATAACCCTCTATTGCAACCATAGAAACAGATCCA AAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCC CACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAAC TGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAG AGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTC TATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAG GAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTAC GAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG

In some embodiments, the polynucleotide of the present invention encodesa CAR, wherein the CAR comprises an amino acid sequence at least about75%, at least about 85%, at least about 85%, at least about 90%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or 100% identical to an amino acid sequenceselected from the group consisting of SEQ ID NOs: 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,178, 180, 190, 192, 202, 204, 214, 216, 226, and 228. In certainembodiments, the CAR comprises an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 134, 136, 138, 140, 142, 144, 146, 148,150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 178, 180, 190, 192,202, 204, 214, 216, 226, and 228.

In some embodiments, the polynucleotide of the present inventioncomprises an nucleotide sequence at least about 50%, at least about 60%,at least about 65%, at least about 70%, at least about 75%, at leastabout 85%, at least about 85%, at least about 90%, at least about 95%,at least about 96%, at least about 97%, at least about 98%, at leastabout 99%, or 100% identical to an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 133, 135, 137, 139, 141, 143, 145, 147,149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 177, 179, 189, 191,201, 203, 213, 215, 225, and 227. In certain embodiments, thepolynucleotide comprises a nucleotide sequence selected from the groupconsisting of SEQ ID NOs: 133, 135, 137, 139, 141, 143, 145, 147, 149,151, 153, 155, 157, 159, 161, 163, 165, 167, 177, 179, 189, 191, 201,203, 213, 215, 225, and 227.

II. Vectors, Cells, and Pharmaceutical Compositions

In certain aspects, provided herein are vectors comprising apolynucleotide of the present invention. In some embodiments, thepresent invention is directed to a vector or a set of vectors comprisinga polynucleotide encoding a CAR or a TCR comprising the truncated hingedomain (“THD”) domain, as described above.

Any vector known in the art can be suitable for the present invention.In some embodiments, the vector is a viral vector. In some embodiments,the vector is a retroviral vector, a DNA vector, a murine leukemia virusvector, an SFG vector, a plasmid, a RNA vector, an adenoviral vector, abaculoviral vector, an Epstein Barr viral vector, a papovaviral vector,a vaccinia viral vector, a herpes simplex viral vector, an adenovirusassociated vector (AAV), a lentiviral vector, or any combinationthereof.

In an embodiment, a vector that can be employed in the context of thepresent invention is pGAR and has the coding sequence:

CTGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTGTAATACGACTCACTATAGGGCGACCCGGGGATGGCGCGCCAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGCTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTGGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCCGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTAACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAATTCAAAATTTTATCGCGATCGCGGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAATACATAACTGAGAATAGAGAAGTTCAGATCAAGGTTAGGAACAGAGAGACAGCAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGCGCGCCAGTCCTTCGAAGTAGATCTTTGTCGATCCTACCATCCACTCGACACACCCGCCAGCGGCCGCTGCCAAGCTTCCGAGCTCTCGAATTAATTCACGGTACCCACCATGGCCTAGGGAGACTAGTCGAATCGATATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTTCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGGTTAATTAAAGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCGAATTCACTCCCAACGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGGCATGCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTGGCGCGCCATCGTCGAGGTTCCCTTTAGTGAGGGTTAATTGCGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCC GAAAAGTGCCAC

The pGAR vector map is set forth below:

Suitable additional exemplary vectors include e.g., pBABE-puro,pBABE-neo largeTcDNA, pBABE-hygro-hTERT, pMKO.1 GFP, MSCV-IRES-GFP,pMSCV PIG (Puro IRES GFP empty plasmid),pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE, MSCV IRES Luciferase, pMIG,MDH1-PGK-GFP_2.0, TtRMPVIR, pMSCV-IRES-mCherry FP, pRetroX GFP T2A Cre,pRXTN, pLncEXP, and pLXIN-Luc.

In other aspects, provided herein are cells comprising a polynucleotideor a vector of the present invention. In some embodiments, the presentinvention is directed to cells, e.g., in vitro cells, comprising apolynucleotide encoding a CAR or a TCR comprising a TCD describedherein. In other embodiments, the present invention is directed tocells, e.g., in vitro cells, comprising a polypeptide encoded by a CARor a TCR comprising a TCD described herein.

Any cell may be used as a host cell for the polynucleotides, thevectors, or the polypeptides of the present invention. In someembodiments, the cell can be a prokaryotic cell, fungal cell, yeastcell, or higher eukaryotic cells such as a mammalian cell. Suitableprokaryotic cells include, without limitation, eubacteria, such asGram-negative or Gram-positive organisms, for example, Enterobactehaceaesuch as Escherichia, e.g., E. coli; Enterobacter; Erwinia; Klebsiella;Proteus; Salmonella, e.g., Salmonella typhimurium; Serratia, e.g.,Serratia marcescans, and Shigella; Bacilli such as B. subtilis and B.licheniformis; Pseudomonas such as P. aeruginosa; and Streptomyces. Insome embodiments, the cell is a human cell. In some embodiments, thecell is an immune cell. In some embodiments, the immune cell is selectedfrom the group consisting of a T cell, a B cell, a tumor infiltratinglymphocyte (TIL), a TCR expressing cell, a natural killer (NK) cell, adendritic cell, a granulocyte, an innate lymphoid cell, a megakaryocyte,a monocyte, a macrophage, a platelet, a thymocyte, and a myeloid cell.In one embodiment, the immune cell is a T cell. In another embodiment,the immune cell is an NK cell. In certain embodiments, the T cell is atumor-infiltrating lymphocyte (TIL), autologous T cell, engineeredautologous T cell (eACT™), an allogeneic T cell, a heterologous T cell,or any combination thereof.

The cell of the present invention may be obtained through any sourceknown in the art. For example, T cells can be differentiated in vitrofrom a hematopoietic stem cell population, or T cells can be obtainedfrom a subject. T cells can be obtained from, e.g., peripheral bloodmononuclear cells, bone marrow, lymph node tissue, cord blood, thymustissue, tissue from a site of infection, ascites, pleural effusion,spleen tissue, and tumors. In addition, the T cells can be derived fromone or more T cell lines available in the art. T cells can also beobtained from a unit of blood collected from a subject using any numberof techniques known to the skilled artisan, such as FICOLL™ separationand/or apheresis. In certain embodiments, the cells collected byapheresis are washed to remove the plasma fraction, and placed in anappropriate buffer or media for subsequent processing. In someembodiments, the cells are washed with PBS. As will be appreciated, awashing step can be used, such as by using a semiautomated flowthroughcentrifuge, e.g., the Cobe™ 2991 cell processor, the Baxter CytoMate™,or the like. In some embodiments, the washed cells are resuspended inone or more biocompatible buffers, or other saline solution with orwithout buffer. In certain embodiments, the undesired components of theapheresis sample are removed. Additional methods of isolating T cellsfor a T cell therapy are disclosed in U.S. Patent Publication No.2013/0287748, which is herein incorporated by references in itsentirety.

In certain embodiments, T cells are isolated from PBMCs by lysing thered blood cells and depleting the monocytes, e.g., by usingcentrifugation through a PERCOLL™ gradient. In some embodiments, aspecific subpopulation of T cells, such as CD4⁺, CD8⁺, CD28⁺, CD45RA⁺,and CD45RO⁺ T cells is further isolated by positive or negativeselection techniques known in the art. For example, enrichment of a Tcell population by negative selection can be accomplished with acombination of antibodies directed to surface markers unique to thenegatively selected cells. In some embodiments, cell sorting and/orselection via negative magnetic immunoadherence or flow cytometry thatuses a cocktail of monoclonal antibodies directed to cell surfacemarkers present on the cells negatively selected can be used. Forexample, to enrich for CD4⁺ cells by negative selection, a monoclonalantibody cocktail typically includes antibodies to CD8, CD11b, CD14,CD16, CD20, and HLA-DR. In certain embodiments, flow cytometry and cellsorting are used to isolate cell populations of interest for use in thepresent invention.

In some embodiments, PBMCs are used directly for genetic modificationwith the immune cells (such as CARs or TCRs) using methods as describedherein. In certain embodiments, after isolating the PBMCs, T lymphocytesare further isolated, and both cytotoxic and helper T lymphocytes aresorted into naive, memory, and effector T cell subpopulations eitherbefore or after genetic modification and/or expansion.

In some embodiments, CD8⁺ cells are further sorted into naive, centralmemory, and effector cells by identifying cell surface antigens that areassociated with each of these types of CD8⁺ cells. In some embodiments,the expression of phenotypic markers of central memory T cells includesCCR7, CD3, CD28, CD45RO, CD62L, and CD127 and are negative for granzymeB. In some embodiments, central memory T cells are CD8⁺, CD45RO⁺, andCD62L⁺ T cells. In some embodiments, effector T cells are negative forCCR7, CD28, CD62L, and CD127 and positive for granzyme B and perforin.In certain embodiments, CD4⁺ T cells are further sorted intosubpopulations. For example, CD4⁺ T helper cells can be sorted intonaive, central memory, and effector cells by identifying cellpopulations that have cell surface antigens.

In some embodiments, the immune cells, e.g., T cells, are geneticallymodified following isolation using known methods, or the immune cellsare activated and expanded (or differentiated in the case ofprogenitors) in vitro prior to being genetically modified. In anotherembodiment, the immune cells, e.g., T cells, are genetically modifiedwith the chimeric antigen receptors described herein (e.g., transducedwith a viral vector comprising one or more nucleotide sequences encodinga CAR) and then are activated and/or expanded in vitro. Methods foractivating and expanding T cells are known in the art and are described,e.g., in U.S. Pat. Nos. 6,905,874; 6,867,041; and 6,797,514; and PCTPublication No. WO 2012/079000, the contents of which are herebyincorporated by reference in their entirety. Generally, such methodsinclude contacting PBMC or isolated T cells with a stimulatory agent andcostimulatory agent, such as anti-CD3 and anti-CD28 antibodies,generally attached to a bead or other surface, in a culture medium withappropriate cytokines, such as IL-2. Anti-CD3 and anti-CD28 antibodiesattached to the same bead serve as a “surrogate” antigen presenting cell(APC). One example is The Dynabeads® system, a CD3/CD28activator/stimulator system for physiological activation of human Tcells. In other embodiments, the T cells are activated and stimulated toproliferate with feeder cells and appropriate antibodies and cytokinesusing methods such as those described in U.S. Pat. Nos. 6,040,177 and5,827,642 and PCT Publication No. WO 2012/129514, the contents of whichare hereby incorporated by reference in their entirety.

In certain embodiments, the T cells are obtained from a donor subject.In some embodiments, the donor subject is human patient afflicted with acancer or a tumor. In other embodiments, the donor subject is a humanpatient not afflicted with a cancer or a tumor.

Other aspects of the present invention are directed to compositionscomprising a polynucleotide described herein, a vector described herein,a polypeptide described herein, or an in vitro cell described herein. Insome embodiments, the composition comprises a pharmaceuticallyacceptable carrier, diluent, solubilizer, emulsifier, preservativeand/or adjuvant. In some embodiments, the composition comprises anexcipient. In one embodiment, the composition comprises a polynucleotideencoding a CAR or a TCR comprising a truncated hinge domain (“THD”)described herein. In another embodiment, the composition comprises a CARor a TCR comprising a TCD encoded by a polynucleotide of the presentinvention. In another embodiment, the composition comprises a T cellcomprising a CAR or a TCR comprising a TCD described herein.

In other embodiments, the composition is selected for parenteraldelivery, for inhalation, or for delivery through the digestive tract,such as orally. The preparation of such pharmaceutically acceptablecompositions is within the ability of one skilled in the art. In certainembodiments, buffers are used to maintain the composition atphysiological pH or at a slightly lower pH, typically within a pH rangeof from about 5 to about 8. In certain embodiments, when parenteraladministration is contemplated, the composition is in the form of apyrogen-free, parenterally acceptable aqueous solution comprising acomposition described herein, with or without additional therapeuticagents, in a pharmaceutically acceptable vehicle. In certainembodiments, the vehicle for parenteral injection is sterile distilledwater in which composition described herein, with or without at leastone additional therapeutic agent, is formulated as a sterile, isotonicsolution, properly preserved. In certain embodiments, the preparationinvolves the formulation of the desired molecule with polymericcompounds (such as polylactic acid or polyglycolic acid), beads orliposomes, that provide for the controlled or sustained release of theproduct, which are then be delivered via a depot injection. In certainembodiments, implantable drug delivery devices are used to introduce thedesired molecule.

III. Methods of the Invention

Another aspect of the invention is directed to a method of making a cellexpressing a CAR or a TCR comprising transducing a cell with apolynucleotide disclosed herein under suitable conditions. In someembodiments, the method comprises transducing a cell with apolynucleotide encoding a CAR or a TCR, as disclosed herein. In someembodiments, the method comprises transducing a cell with a vectorcomprising the polynucleotide encoding a CAR or a TCR.

Another aspect of the present invention is directed to a method ofinducing an immunity against a tumor comprising administering to asubject an effective amount of a cell comprising a polynucleotidedescribed herein, a vector described herein, or a CAR or a TCR describedherein. In one embodiment, the method comprises administering to asubject an effective amount of a cell comprising a polynucleotideencoding a CAR or a TCR disclosed herein. In another embodiment, themethod comprises administering to a subject an effective amount of acell comprising a vector comprising a polynucleotide encoding a CAR or aTCR disclosed herein. In another embodiment, the method comprisesadministering to a subject an effective amount of a cell comprising aCAR or a TCR encoded by a polynucleotide disclosed herein.

Another aspect of the present invention is directed to a method ofinducing an immune response in a subject comprising administering aneffective amount of the engineered immune cells of the presentapplication. In some embodiments, the immune response is a Tcell-mediated immune response. In some embodiments, the T cell-mediatedimmune response is directed against one or more target cells. In someembodiments, the engineered immune cell comprises a CAR or a TCR,wherein the CAR or the TCR comprises a THD described in the presentdisclosure. In some embodiments, the target cell is a tumor cell.

Another aspect of the present invention is directed to a method fortreating or preventing a malignancy, said method comprisingadministering to a subject in need thereof an effective amount of atleast one immune cell, wherein the immune cell comprises at least oneCAR or TCR, and wherein the CAR or the TCR comprises a THD describedherein.

Another aspect of the present invention is directed to a method oftreating a cancer in a subject in need thereof comprising administeringto the subject a polynucleotide, a vector, a CAR or a TCR, a cell, or acomposition disclosed herein. In one embodiment, the method comprisesadministering a polynucleotide encoding a CAR or a TCR. In anotherembodiment, the method comprises administering a vector comprising apolynucleotide encoding a CAR or a TCR. In another embodiment, themethod comprises administering a CAR or a TCR encoded by apolynucleotide disclosed herein. In another embodiment, the methodcomprises administering a cell comprising the polynucleotide, or avector comprising the polynucleotide, encoding a CAR or a TCR.

In some embodiments, the methods of treating a cancer in a subject inneed thereof comprise a T cell therapy. In one embodiment, the T celltherapy of the present invention is engineered Autologous Cell Therapy(eACT™). According to this embodiment, the method can include collectingblood cells from the patient. The isolated blood cells (e.g., T cells)can then be engineered to express a CAR or a TCR of the presentinvention. In a particular embodiment, the CAR T cells or the TCR Tcells are administered to the patient. In some embodiments, the CAR Tcells or the TCR T cells treat a tumor or a cancer in the patient. Inone embodiment the CAR T cells or the TCR T cells reduce the size of atumor or a cancer.

In some embodiments, the donor T cells for use in the T cell therapy areobtained from the patient (e.g., for an autologous T cell therapy). Inother embodiments, the donor T cells for use in the T cell therapy areobtained from a subject that is not the patient.

The T cells can be administered at a therapeutically effective amount.For example, a therapeutically effective amount of the T cells can be atleast about 10⁴ cells, at least about 10⁵ cells, at least about 10⁶cells, at least about 10⁷ cells, at least about 10⁸ cells, at leastabout 10⁹, or at least about 10¹⁰. In another embodiment, thetherapeutically effective amount of the T cells is about 10⁴ cells,about 10⁵ cells, about 10⁶ cells, about 10⁷ cells, or about 10⁸ cells.In one particular embodiment, the therapeutically effective amount ofthe CAR T cells or the TCR T cells is about 2×10⁶ cells/kg, about 3×10⁶cells/kg, about 4×10⁶ cells/kg, about 5×10⁶ cells/kg, about 6×10⁶cells/kg, about 7×10⁶ cells/kg, about 8×10⁶ cells/kg, about 9×10⁶cells/kg, about 1×10⁷ cells/kg, about 2×10⁷ cells/kg, about 3×10⁷cells/kg, about 4×10⁷ cells/kg, about 5×10⁷ cells/kg, about 6×10⁷cells/kg, about 7×10⁷ cells/kg, about 8×10⁷ cells/kg, or about 9×10⁷cells/kg.

IV. Cancer Treatment

The methods of the invention can be used to treat a cancer in a subject,reduce the size of a tumor, kill tumor cells, prevent tumor cellproliferation, prevent growth of a tumor, eliminate a tumor from apatient, prevent relapse of a tumor, prevent tumor metastasis, induceremission in a patient, or any combination thereof. In certainembodiments, the methods induce a complete response. In otherembodiments, the methods induce a partial response.

Cancers that may be treated include tumors that are not vascularized,not yet substantially vascularized, or vascularized. The cancer may alsoinclude solid or non-solid tumors. In some embodiments, the cancer is ahematologic cancer. In some embodiments, the cancer is of the whiteblood cells. In other embodiments, the cancer is of the plasma cells. Insome embodiments, the cancer is leukemia, lymphoma, or myeloma. Incertain embodiments, the cancer is acute lymphoblastic leukemia (ALL)(including non T cell ALL), acute lymphoid leukemia (ALL), andhemophagocytic lymphohistocytosis (HLH)), B cell prolymphocyticleukemia, B-cell acute lymphoid leukemia (“BALL”), blastic plasmacytoiddendritic cell neoplasm, Burkitt's lymphoma, chronic lymphocyticleukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloidleukemia (CIVIL), chronic or acute granulomatous disease, chronic oracute leukemia, diffuse large B cell lymphoma, diffuse large B celllymphoma (DLBCL), follicular lymphoma, follicular lymphoma (FL), hairycell leukemia, hemophagocytic syndrome (Macrophage Activating Syndrome(MAS), Hodgkin's Disease, large cell granuloma, leukocyte adhesiondeficiency, malignant lymphoproliferative conditions, MALT lymphoma,mantle cell lymphoma, Marginal zone lymphoma, monoclonal gammapathy ofundetermined significance (MGUS), multiple myeloma, myelodysplasia andmyelodysplastic syndrome (MDS), myeloid diseases including but notlimited to acute myeloid leukemia (AML), non-Hodgkin's lymphoma (NHL),plasma cell proliferative disorders (e.g., asymptomatic myeloma(smoldering multiple myeloma or indolent myeloma), plasmablasticlymphoma, plasmacytoid dendritic cell neoplasm, plasmacytomas (e.g.,plasma cell dyscrasia; solitary myeloma; solitary plasmacytoma;extramedullary plasmacytoma; and multiple plasmacytoma), POEMS syndrome(Crow-Fukase syndrome; Takatsuki disease; PEP syndrome), primarymediastinal large B cell lymphoma (PMBC), small cell- or a largecell-follicular lymphoma, splenic marginal zone lymphoma (SMZL),systemic amyloid light chain amyloidosis, T-cell acute lymphoid leukemia(“TALL”), T-cell lymphoma, transformed follicular lymphoma, Waldenstrommacroglobulinemia, or a combination thereof.

In one embodiment, the cancer is a myeloma. In one particularembodiment, the cancer is multiple myeloma. In another embodiment, thecancer is a leukemia. In one embodiment, the cancer is acute myeloidleukemia.

In some embodiments, the methods further comprise administering achemotherapeutic. In certain embodiments, the chemotherapeutic selectedis a lymphodepleting (preconditioning) chemotherapeutic. Beneficialpreconditioning treatment regimens, along with correlative beneficialbiomarkers are described in U.S. Provisional Patent Applications62/262,143 and 62/167,750 which are hereby incorporated by reference intheir entirety herein. These describe, e.g., methods of conditioning apatient in need of a T cell therapy comprising administering to thepatient specified beneficial doses of cyclophosphamide (between 200mg/m²/day and 2000 mg/m²/day) and specified doses of fludarabine(between 20 mg/m²/day and 900 mg/m²/day). One such dose regimen involvestreating a patient comprising administering daily to the patient about500 mg/m²/day of cyclophosphamide and about 60 mg/m²/day of fludarabinefor three days prior to administration of a therapeutically effectiveamount of engineered T cells to the patient.

In other embodiments, the antigen binding molecule, transduced (orotherwise engineered) cells (such as CARs or TCRs), and thechemotherapeutic agent are administered each in an amount effective totreat the disease or condition in the subject.

In certain embodiments, compositions comprising CAR- and/orTCR-expressing immune effector cells disclosed herein may beadministered in conjunction with any number of chemotherapeutic agents.Examples of chemotherapeutic agents include alkylating agents such asthiotepa and cyclophosphamide (CYTOXAN™); alkyl sulfonates such asbusulfan, improsulfan and piposulfan; aziridines such as benzodopa,carboquone, meturedopa, and uredopa; ethylenimines and methylamelaminesincluding altretamine, triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide and trimethylolomelamine resume; nitrogenmustards such as chlorambucil, chlornaphazine, cholophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembichin, phenesterine, prednimustine,trofosfamide, uracil mustard; nitrosureas such as carmustine,chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin,carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine,5-FU; androgens such as calusterone, dromostanolone propionate,epitiostanol, mepitiostane, testolactone; anti-adrenals such asaminoglutethimide, mitotane, trilostane; folic acid replenisher such asfrolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinicacid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine;demecolcine; diaziquone; elformithine; elliptinium acetate; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone;mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane;sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g.paclitaxel (TAXOL™, Bristol-Myers Squibb) and doxetaxel (TAXOTERE®,Rhone-Poulenc Rorer); chlorambucil; gemcitabine; 6-thioguanine;mercaptopurine; methotrexate; platinum analogs such as cisplatin andcarboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide;mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine;novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate;CPT-11; topoisomerase inhibitor RFS2000; difluoromethylomithine (DMFO);retinoic acid derivatives such as Targretin™ (bexarotene), Panretin™,(alitretinoin); ONTAK™ (denileukin diftitox); esperamicins;capecitabine; and pharmaceutically acceptable salts, acids orderivatives of any of the above. In some embodiments, compositionscomprising CAR- and/or TCR-expressing immune effector cells disclosedherein may be administered in conjunction with an anti-hormonal agentthat acts to regulate or inhibit hormone action on tumors such asanti-estrogens including for example tamoxifen, raloxifene, aromataseinhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene,LY117018, onapristone, and toremifene (Fareston); and anti-androgenssuch as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin;and pharmaceutically acceptable salts, acids or derivatives of any ofthe above. Combinations of chemotherapeutic agents are also administeredwhere appropriate, including, but not limited to CHOP, i.e.,Cyclophosphamide (Cytoxan®), Doxorubicin (hydroxydoxorubicin),Vincristine (Oncovin®), and Prednisone.

In some embodiments, the chemotherapeutic agent is administered at thesame time or within one week after the administration of the engineeredcell or nucleic acid. In other embodiments, the chemotherapeutic agentis administered from 1 to 4 weeks or from 1 week to 1 month, 1 week to 2months, 1 week to 3 months, 1 week to 6 months, 1 week to 9 months, or 1week to 12 months after the administration of the engineered cell ornucleic acid. In some embodiments, the chemotherapeutic agent isadministered at least 1 month before administering the cell or nucleicacid. In some embodiments, the methods further comprise administeringtwo or more chemotherapeutic agents.

A variety of additional therapeutic agents may be used in conjunctionwith the compositions described herein. For example, potentially usefuladditional therapeutic agents include PD-1 inhibitors such as nivolumab(OPDIVO®), pembrolizumab (KEYTRUDA®), pembrolizumab, pidilizumab(CureTech), and atezolizumab (Roche).

Additional therapeutic agents suitable for use in combination with theinvention include, but are not limited to, ibrutinib (IMBRUVICA®),ofatumumab (ARZERRA®), rituximab (RITUXAN®), bevacizumab (AVASTIN®),trastuzumab (HERCEPTIN®), trastuzumab emtansine (KADCYLA®), imatinib(GLEEVEC®), cetuximab (ERBITUX®), panitumumab (VECTIBIX®), catumaxomab,ibritumomab, ofatumumab, tositumomab, brentuximab, alemtuzumab,gemtuzumab, erlotinib, gefitinib, vandetanib, afatinib, lapatinib,neratinib, axitinib, masitinib, pazopanib, sunitinib, sorafenib,toceranib, lestaurtinib, axitinib, cediranib, lenvatinib, nintedanib,pazopanib, regorafenib, semaxanib, sorafenib, sunitinib, tivozanib,toceranib, vandetanib, entrectinib, cabozantinib, imatinib, dasatinib,nilotinib, ponatinib, radotinib, bosutinib, lestaurtinib, ruxolitinib,pacritinib, cobimetinib, selumetinib, trametinib, binimetinib,alectinib, ceritinib, crizotinib, aflibercept, adipotide, denileukindiftitox, mTOR inhibitors such as Everolimus and Temsirolimus, hedgehoginhibitors such as sonidegib and vismodegib, CDK inhibitors such as CDKinhibitor (palbociclib).

In additional embodiments, the composition comprising CAR- and/orTCR-containing immune are administered with an anti-inflammatory agent.Anti-inflammatory agents or drugs can include, but are not limited to,steroids and glucocorticoids (including betamethasone, budesonide,dexamethasone, hydrocortisone acetate, hydrocortisone, hydrocortisone,methylprednisolone, prednisolone, prednisone, triamcinolone),nonsteroidal anti-inflammatory drugs (NSAIDS) including aspirin,ibuprofen, naproxen, methotrexate, sulfasalazine, leflunomide, anti-TNFmedications, cyclophosphamide and mycophenolate. Exemplary NSAIDsinclude ibuprofen, naproxen, naproxen sodium, Cox-2 inhibitors, andsialylates. Exemplary analgesics include acetaminophen, oxycodone,tramadol of proporxyphene hydrochloride. Exemplary glucocorticoidsinclude cortisone, dexamethasone, hydrocortisone, methylprednisolone,prednisolone, or prednisone. Exemplary biological response modifiersinclude molecules directed against cell surface markers (e.g., CD4, CD5,etc.), cytokine inhibitors, such as the TNF antagonists, (e.g.,etanercept (ENBREL®), adalimumab (HUMIRA®) and infliximab (REMICADE®),chemokine inhibitors and adhesion molecule inhibitors. The biologicalresponse modifiers include monoclonal antibodies as well as recombinantforms of molecules. Exemplary DMARDs include azathioprine,cyclophosphamide, cyclosporine, methotrexate, penicillamine,leflunomide, sulfasalazine, hydroxychloroquine, Gold (oral (auranofin)and intramuscular), and minocycline.

In certain embodiments, the compositions described herein areadministered in conjunction with a cytokine. “Cytokine” as used hereinis meant to refer to proteins released by one cell population that acton another cell as intercellular mediators. Examples of cytokines arelymphokines, monokines, and traditional polypeptide hormones. Includedamong the cytokines are growth hormones such as human growth hormone,N-methionyl human growth hormone, and bovine growth hormone; parathyroidhormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin;glycoprotein hormones such as follicle stimulating hormone (FSH),thyroid stimulating hormone (TSH), and luteinizing hormone (LH); hepaticgrowth factor (HGF); fibroblast growth factor (FGF); prolactin;placental lactogen; mullerian-inhibiting substance; mousegonadotropin-associated peptide; inhibin; activin; vascular endothelialgrowth factor; integrin; thrombopoietin (TPO); nerve growth factors(NGFs) such as NGF-beta; platelet-growth factor; transforming growthfactors (TGFs) such as TGF-alpha and TGF-beta; insulin-like growthfactor-I and -II; erythropoietin (EPO); osteoinductive factors;interferons such as interferon-alpha, beta, and -gamma; colonystimulating factors (CSFs) such as macrophage-CSF (M-CSF);granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (G-CSF);interleukins (ILs) such as IL-1, IL-1alpha, IL-2, IL-3, IL-4, IL-5,IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; IL-15, a tumor necrosisfactor such as TNF-alpha or TNF-beta; and other polypeptide factorsincluding LIF and kit ligand (KL). As used herein, the term cytokineincludes proteins from natural sources or from recombinant cell culture,and biologically active equivalents of the native sequence cytokines.

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.However, the citation of a reference herein should not be construed asan acknowledgement that such reference is prior art to the presentinvention. To the extent that any of the definitions or terms providedin the references incorporated by reference differ from the terms anddiscussion provided herein, the present terms and definitions control.

The present invention is further illustrated by the following exampleswhich should not be construed as further limiting. The contents of allreferences cited throughout this application are expressly incorporatedherein by reference.

EXAMPLES Example 1

Plasmids encoding a T7 promoter, CAR construct and a beta globinstabilizing sequence were linearized by overnight digestion of 10 μg DNAwith EcoRI and BamHI (NEB). DNA was then digested for 2 hours at 50° C.with proteinase K (Thermo Fisher, 600 U/ml) purified withphenol/chloroform and precipitated by adding sodium acetate and twovolumes of ethanol. Pellets were then dried, resuspended inRNAse/DNAse-free water and quantified using NanoDrop. One μg of thelinear DNA was then used for in vitro transcription using the mMESSAGEmMACHINE T7 Ultra (Thermo Fisher) following the manufacturer'sinstructions. RNA was further purified using the MEGAClear Kit (ThermoFisher) following the manufacturer's instructions and quantified usingNanoDrop. mRNA integrity was assessed using mobility on an agarose gel.PBMCs were isolated from healthy donor leukopaks (Hemacare) usingficoll-paque density centrifugation per manufacturer's instructions.PBMCs were stimulated using OKT3 (50 ng/ml, Miltenyi Biotec) in R10medium+IL-2 (300 IU/ml, Proleukin®, Prometheus® Therapeutics andDiagnostics). Seven days post-stimulation, T cells were washed twice inOpti-MEM medium (Thermo Fisher Scientific) and resuspended at a finalconcentration of 2.5×10⁷ cells/ml in Opti-MEM medium. Ten μg of mRNA wasused per electroporation. Electroporation of cells was performed using aGemini X2 system (Harvard Apparatus BTX) to deliver a single 400 V pulsefor 0.5 ms in 2 mm cuvettes (Harvard Apparatus BTX). Cells wereimmediately transferred to R10+IL-2 medium and allowed to recover for 6hours. To examine CAR expression, T cells were stained with FLT-=3-HIS(Sino Biological Inc.) or biotinylated Protein L (Thermo Scientific) instain buffer (BD Pharmingen) for 30 minutes at 4° C. Cells were thenwashed and stained with anti-HIS-PE (Miltenyi Biotec) or PE Streptavidin(BD Pharmingen) in stain buffer for 30 minutes at 4° C. Cells were thenwashed and resuspended in stain buffer with propidium iodide (BDPharmingen) prior to data acquisition. Expression of FLT3 CARs inelectroporated T cells is shown in FIG. 3.

T cells were electroporated with plasmids encoding an anti-FLT3 CARcomprising a 10E3, 2E7, 8B5, 4E9, or 11F11 anti-FLT3 binding moleculeand a hinge region selected from the full length hinge domain (acomplete hinge domain or “CHD”) or a truncated hinge domain (“THD”). Theelectroporated anti-FLT3 CAR T cells were then co-cultured with Namalwa(FLT3 negative), EoL1 (FLT3 positive), HL60 (FLT3 positive), or MV4;11(FLT3 positive) target cells at a 1:1 E:T ratio in R10 medium. Sixteenhours post-co-culture, supernatants from Namalwa (FIGS. 4A-4F), EoL1(FIGS. 4G-4L), HL60 (FIGS. 4M-4R, and MV4;11 (4S-4X) were analyzed byLuminex (EMD Millipore) for production of IFNγ (FIGS. 4A, 4B, 4G, 4H,4M, 4N, 4S, and 4T), IL-2 (FIGS. 4C, 4D, 4I, 4J, 40, 4P, 4U, and 4V),and TNFα (FIGS. 4E, 4F, 4K, 4L, 4Q, 4R, 4W, and 4X).

Target cell viability was assessed by flow cytometric analysis ofpropidium iodide (PI) uptake by CD3-negative cells. The electroporatedanti-FLT3 CAR T cells were co-cultured with Namalwa (FIGS. 5A-5B), EoL1(FIGS. 5C-5D), HL60 (FIGS. 5E-5F, and MV4;11 (5G-5H) target cells at 16hours post-co-culture.

Example 2

A third generation lentiviral transfer vector containing the differentCAR constructs was used along with the ViraPower Lentiviral PackagingMix (Life Technologies) to generate the lentiviral supernatants.Briefly, a transfection mix was generated by mixing 15 of DNA and 22.5μl of polyethileneimine (Polysciences, 1 mg/ml) in 600 μl of OptiMEMmedium. The mix was incubated for 5 minutes at room temperature.Simultaneously, 293T cells (ATCC) were trypsinized, counted and a totalof 10×10⁶ total cells were plated in a T75 flask along the transfectionmix. Three days after the transfection, supernatants were collected andfiltered through a 0.45 μm filter and stored at −80° C. until used.PBMCs were isolated from healthy donor leukopaks (Hemacare) usingficoll-paque density centrifugation per manufacturer's instructions.PBMCs were stimulated using OKT3 (50 ng/ml, Miltenyi Biotec) in R10medium+IL-2 (300 IU/ml, PROLEUKIN®, PROMETHEUS® Therapeutics andDiagnostics). Forty eight hours post-stimulation, cells were transducedusing lentivirus at an MOI=10. Cells were maintained at 0.5-2.0×10⁶cells/ml prior to use in activity assays. To examine CAR expression, Tcells were stained with FLT-3-HIS (Sino Biological Inc.) or biotinylatedProtein L (Thermo Scientific) in stain buffer (BD Pharmingen) for 30minutes at 4° C. Cells were then washed and stained with anti-HIS-PE(Miltenyi Biotec) or PE Streptavidin (BD Pharmingen) in stain buffer for30 minutes at 4° C. Cells were then washed and resuspended in stainbuffer with propidium iodide (BD Pharmingen) prior to data acquisition.Expression of FLT3 CARs in T cells from two healthy donors is shown inFIG. 6A-6B.

T cells from two healthy donors were transduced with lentiviral vectorsencoding anti-FLT3 CAR T cells comprising a 10E3, 8B5, or 11F11 bindingmolecule and a hinge region selected from the complete hinge domain(“CHD”), a truncated hinge domain (“THD”), and the CD8 hinge region.Transduced T cells were co-cultured with target cells at a 1:1 E:T ratioin R10 medium. Sixteen hours post-co-culture, supernatants were analyzedby Luminex (EMD Millipore) for production of IFNγ (FIGS. 7A-7B), TNFα(FIGS. 7C-7D), and IL-2 (FIGS. 7E-7F).

Target cell viability was assessed by flow cytometric analysis ofpropidium iodide (PI) uptake by CD3-negative cells. Average cytolyticactivity of lentivirus-transduced CAR T cells (from two healthy donors)co-cultured with Namalwa (FIG. 8A), EoL1 (FIG. 8B), MV4;11 (FIG. 8C),and HL60 (FIG. 8D) target cells was measured.

To assess CAR T cell proliferation in response to FLT3 expressing targetcells, T cells were labeled with CFSE prior to co-culture with targetcells at a 1:1 E:T ratio in R10 medium. Five days later, T cellproliferation was assessed by flow cytometric analysis of CFSE dilution.Proliferation of FLT3 CAR T cells is shown in FIGS. 9A-9B.

Example 3

To examine in vivo anti-leukemic activity, FLT3 CAR T cells weregenerated for use in a xenogeneic model of human AML. CAR expression ofthe various effector lines used in the xenogeneic model of human AML areshown in FIGS. 10A-10D. Luciferase-labeled MV4;11 cells (2×10⁶cells/animal) were injected intravenously into 5 to 6 week-old femaleNSG mice. After 6 days, 6×10⁶ T cells (˜50% CAR+) in 200 μl PBS wereinjected intravenously, and the tumor burden of the animals was measuredweekly using bioluminescence imaging (FIGS. 10E-10G). Survival analysiswas performed by injection of controls (mock) or 10E3-CHD (FIG. 10H),10E3-THD (FIG. 10I), or 8B5-THD (FIG. 10J) expressing CAR T cells.

Example 4

T cells were electroporated with plasmids encoding the anti-CLL-1 CARconstructs 24C8_HL-CHD CAR (comprising a complete hinge domain of thecostimulatory protein) and 24C8_HL-THD CAR (comprising a truncated hingedomain of the costimulatory protein). Anti-CLL-1 expression byelectroporated T cells is shown in FIGS. 11A-11D. The anti-CLL-1 CARTcells were then cultured with the target Namalwa (ATCC; CLL-1 negative),U937 (ATCC; CLL-1 positive), HL-60 (ATCC; CLL-1 positive), EoL-1 (Sigma;CLL-1 positive), KG1a (ATCC; CLL-1 positive) and MV4;11 (ATCC; CLL-1positive) cells at a 1:1 E:T ratio in R10 media 6 hours after mRNAelectroporation. Sixteen hours post-co-culture, supernatants wereanalyzed by Luminex (EMD Millipore), according to the manufacturer'sinstructions, for production of IL-2 (FIG. 12A), IFNγ (FIG. 12B), andTNFα (FIG. 12C).

Target cell viability was assessed by flow cytometric analysis ofpropidium iodide (PI) uptake. The electroporated anti-CLL-1 CAR T cellswere co-cultured with Namalwa (FIG. 13A), MV4;11 (FIG. 13B), EoL-1 (FIG.13C), HL-60 (FIG. 13D), or U937 (FIG. 13E) target cells for 16 hours. Asexpected, Namalwa cells co-cultured with the anti-CLL-1 CART cellsshowed little change in target cell viability, relative to controls(FIG. 13A). However, increased cytolytic activity was observed in MV;411cells co-cultured with 24C8_HL-CHD and 24C8_HL-THD T cells, relative tocontrols, with a greater target cell cytolytic activity observed in the24C8_HL-THD T cell co-culture (FIG. 13B). In addition, increasedcytolytic activity was observed in EoL-1 cells co-cultured with24C8_HL-CHD and 24C8_HL-THD T cells, relative to controls (FIG. 13C).Increased cytolytic activity was observed in HL-60 cells co-culturedwith 24C8_HL-CHD and 24C8_HL-THD T cells, relative to controls (FIG.13D). Increased cytolytic activity was observed in U937 cellsco-cultured with 24C8_HL-CHD and 24C8_HL-THD T cells, relative tocontrols, with a greater target cell cytolytic activity observed in the24C8_HL-THD T cell co-culture (FIG. 13E).

Example 5

T cells transduced with lentiviral vectors comprising an anti-CLL-1 CARconstruct with a truncated hinge domain (“THD”) of the costimulatoryprotein, 10E3 THD or 24C1_LH_THD, were co-cultured with Namalwa, U937,HL-60, EoL-1, KG1a and MV4;11 target cells at a 1:1 E:T ratio in R10media 12 days after T cell stimulation. Sixteen hours post-co-culture,supernatants were analyzed by Luminex (EMD Millipore), according to themanufacturer's instructions, for production of the cytokines IFNγ (FIG.14A), IL-2 (FIG. 14B), and TNFα (FIG. 14C) in co-cultures of effector10E3 THD CART cells and 24C1_LH_THD CAR T cells with target Namalwa,HL-60, or MVA;11 cells, as indicated.

Target cell viability was assessed by flow cytometric analysis ofpropidium iodide (PI) uptake. Transduced effector 24C1_LH_THD CAR Tcells were co-cultured with Namalwa, U937, HL-60, EoL-1, KG1a, or MV4;11target cells for 16 hours or 40 hours. Co-culture of Namalwa targetcells with transduced C1_24C1_LH_THD CART cells had no effect on thepercent of viable Namalwa target cells at 16 hours and 40 hours, ascompared to mock controls (FIG. 15A). However, C1_24C1_LH_THD CAR Tcells co-cultured with either MV4;11 (FIG. 15B) or HL-60 (FIG. 15C)target cells resulted in a lower percent of viable target cells at both16 hours and 40 hours, as compared to mock controls.

Example 6

CAR T cells transduced with anti-BCMA CAR constructs comprising atruncated hinge domain (“THD”) of the costimulatory protein werecultured with target cells at a 1:1 effector cell to target cell (E:T)ratio in R10 media 12 days after T cell stimulation. Cell lines testedincluded EoL-1 (Sigma; BCMA negative), NCI-H929 (Molecular Imaging; BCMApositive), and MM1S (Molecular Imaging; BCMA positive). Sixteen hourspost-co-culture, supernatants were analyzed by Luminex (EMD Millipore),according to the manufacturer's instructions, for production of thecytokines IFNγ (FIGS. 16A-16B), TNFα (FIGS. 16C-16D), and IL-2 (FIGS.16E-16F). IFNγ (FIGS. 16A-16B), TNFα (FIGS. 16C-16D), and IL-2 (FIGS.16E-16F) were observed in the supernatant of NCI-H929 and MM1S targetcell co-cultures for each anti-BCMA CAR T cell tested in both donors(FIGS. 16A-16B); however, IFNγ (FIGS. 16A-16B), TNFα (FIGS. 16C-16D),and IL-2 (FIGS. 16E-16F) were only observed in the supernatant of EoL-1target cell co-cultures above background for the IR negative control Tcells (FIG. 16A).

Target cell viability was assessed by flow cytometric analysis ofpropidium iodide (PI) uptake of CD3 negative cells. The anti-BCMA CAR Tcells were co-cultured with EoL1 (FIGS. 17A-17B), NCI-H929 (FIGS.17C-17D), or MM1S (FIGS. 17E-17F) target cells for 16 hours, 40 hours,64 hours, 88 hours, or 112 hours. Little cytolytic activity was observedin the EoL-1 co-cultures at any time period for the anti-BCMA CAR Tcells (FIG. 17A-17B). However, co-culture of the anti-BCMA CAR T cellsand the NCI-H929 or MM1S target cells resulted in a decrease in thepercentage of viable target cells at each time point measured for eachof the anti-BCMA CAR T cells.

To examine proliferation, anti-BCMA CAR T cells were labeled withcarboxyfluorescein succinimidyl ester (CFSE) prior to co-culture withEoL-1, NCI-H929, or MM1S target cells at a 1:1 E:T ratio in R10 media.Five days later, T cell proliferation was assessed by flow cytometricanalysis of CFSE dilution (FIGS. 18A-18B).

Example 7

Enhanced stability is a desired property of proteins. This is oftenassessed by determining the melting temperature of a protein undervarious conditions. Proteins with a higher melting temperature aregenerally stable for longer times. When a CAR is more thermostable, itmay be functionally active for longer periods of time on the surface ofa cell.

Thermal stability of the CAR extracellular domain (ECD) with the longerhinge domain, i.e., the complete hinge domain (“CHD”) and the thermalstability of the CAR ECD with a truncated hinge domain (“THD”) wasmeasured using a Bio-Rad C1000 thermal cycler, CFx96 Real-Time system.Unfolding of the proteins was monitored using the fluorescent dye SYPROOrange (Invitrogen) which binds to hydrophobic amino acids that becomesolvent exposed as the protein unfolds. A temperature gradient was setup from 25° C. to 95° C. with 1° C./1 minute increments. Each samplecontained 10 μM recombinant CAR ECD protein and 5×SYPRO Orange(Molecular Probes™ SYPRO™ Orange Protein Gel Stain (5,000× Concentratein DMSO)). The assay was performed in PBS with or without 50 mM NaCl.

As shown in FIG. 19A and FIG. 19B, a CAR's ECD which has a THD showsenhanced thermostability compared to a CAR's ECD which has a CHD, e.g.,including the IEVMYPPPY (SEQ ID NO: 250) motif. These method describedin this example is a useful method for testing stability of mRNAencoding a CAR and the CAR itself, because once a T cell has beentransduced with the mRNA encoding a CAR, the transduced T cell willexpress the CAR and the stability of an individual mRNA or proteincannot be readily assessed.

What is claimed is:
 1. An isolated polynucleotide encoding a chimeric antigen receptor (CAR) or a T cell receptor (TCR), which comprises (i) an antigen binding molecule, (ii) a costimulatory domain, and (iii) an activating domain, wherein the costimulatory domain comprises an extracellular domain, a transmembrane domain, and an intracellular domain, wherein the extracellular domain comprises a truncated hinge domain consisting essentially of or consisting of (i) an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to amino acids 123 to 152 of SEQ ID NO: 1 and, optionally, (ii) one to six amino acids.
 2. The polynucleotide of claim 1, wherein the one to six amino acids are heterologous amino acids.
 3. The polynucleotide of claim 1 or 2, wherein the truncated hinge domain consists essentially of or consists of an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to amino acids 123 to 152 of SEQ ID NO:
 1. 4. The polynucleotide of claim 1 or 2, wherein the amino acid sequence is encoded by a nucleotide sequence at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 2. 5. The polynucleotide of claim 1 or 2, wherein the transmembrane domain is a transmembrane domain of 4-1BB/CD137, an alpha chain of a T cell receptor, a beta chain of a T cell receptor, CD3 epsilon, CD4, CD5, CD8 alpha, CD9, CD16, CD19, CD22, CD33, CD37, CD45, CD64, CD80, CD86, CD134, CD137, CD154, or a zeta chain of a T cell receptor, or any combination thereof.
 6. The polynucleotide of claim 1 or 2, wherein the transmembrane domain comprises an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 5. 7. The polynucleotide of claim 6, wherein the transmembrane domain is encoded by a nucleotide sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 4. 8. The polynucleotide of claim 1 or 2, wherein the intracellular domain comprises a signaling region of 4-1BB/CD137, activating NK cell receptors, B7-H3, BAFFR, BLAME (SLAMF8), BTLA, CD100 (SEMA4D), CD103, CD160 (BY55), CD18, CD19, CD19a, CD2, CD247, CD27, CD276 (B7-H3), CD29, CD3 delta, CD3 epsilon, CD3 gamma, CD30, CD4, CD40, CD49a, CD49D, CD49f, CD69, CD7, CD84, CD8alpha, CD8beta, CD96 (Tactile), CD11a, CD11b, CD11c, CD11d, CDS, CEACAM1, CRT AM, cytokine receptors, DAP-10, DNAM1 (CD226), Fc gamma receptor, GADS, GITR, HVEM (LIGHTR), IA4, ICAM-1, ICAM-1, Ig alpha (CD79a), IL2R beta, IL2R gamma, IL7R alpha, Immunoglobulin-like proteins, inducible T cell costimulator (ICOS), integrins, ITGA4, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX, ITGB2, ITGB7, ITGB1, KIRDS2, LAT, LFA-1, LFA-1, a ligand that specifically binds with CD83, LIGHT, LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), LTBR, Ly9 (CD229), lymphocyte function-associated antigen-1 (LFA-1 (CD11a/CD18), MHC class I molecule, NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), OX-40, PAG/Cbp, programmed death-1 (PD-1), PSGL1, SELPLG (CD162), signaling lymphocytic activation molecules (SLAM proteins), SLAM (SLAMF1; CD150; IPO-3), SLAMF4 (CD244; 2B4), SLAMF6 (NTB-A; Ly108), SLAMF7, SLP-76, TNF receptor proteins, TNFR2, a Toll ligand receptor, TRANCE/RANKL, VLA1, or VLA-6, or a combination thereof.
 9. The polynucleotide of claim 1 or 2, wherein the intracellular domain comprises a 4-1BB/CD137 signaling region.
 10. The polynucleotide of claim 1 or 2, wherein the intracellular domain comprises an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 7. 11. The polynucleotide of claim 1 or 2, wherein the intracellular domain comprises an amino acid sequence encoded by a nucleotide sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 6. 12. The polynucleotide of claim 1 or 2, wherein the antigen binding molecule comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises 3 complementarity determining regions (CDRs) and the VL comprises 3 CDRs.
 13. The polynucleotide of claim 1 or 2, wherein the antigen binding molecule specifically binds an antigen selected from the group consisting of 5T4, alphafetoprotein, B cell maturation antigen (BCMA), CA-125, carcinoembryonic antigen, CD19, CD20, CD22, CD23, CD30, CD33, CD56, CD123, CD138, c-Met, CSPG4, C-type lectin-like molecule 1 (CLL-1), EGFRvIII, epithelial tumor antigen, ERBB2, FLT3, folate binding protein, GD2, GD3, HER1-HER2 in combination, HER2-HER3 in combination, HER2/Neu, HERV-K, HIV-1 envelope glycoprotein gp41, HIV-1 envelope glycoprotein gp120, IL-11Ralpha, kappa chain, lambda chain, melanoma-associated antigen, mesothelin, MUC-1, mutated p53, mutated ras, prostate-specific antigen, ROR1, or VEGFR2, or a combination thereof.
 14. The polynucleotide of claim 1 or 2, wherein the antigen binding molecule specifically binds BCMA, CLL-1, or FLT3.
 15. The polynucleotide of claim 1 or 2, wherein the activation domain comprises an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 9 or SEQ ID NO:
 251. 16. The polynucleotide of claim 1 or 2, wherein the activation domain is encoded by a nucleotide sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 8. 17. The polynucleotide of any one of claims 1 to 16, wherein the CAR or TCR further comprises a leader peptide.
 18. The polynucleotide of claim 17, wherein the leader peptide comprises an amino acid sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 11. 19. The polynucleotide of claim 17, wherein the leader peptide is encoded by a nucleotide sequence at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO:
 10. 20. A vector comprising the polynucleotide of any one of claims 1 to
 19. 21. The vector of claim 20, wherein the vector is an adenoviral vector, an adenovirus-associated vector, a DNA vector, a lentiviral vector, a plasmid, a retroviral vector, or an RNA vector, or any combination thereof.
 22. A polypeptide encoded by the polynucleotide of any one of claims 1 to 19 or the vector of claim 20 or
 21. 23. A cell comprising the polynucleotide of any one of claims 1 to 19, the vector of claim 20 or 21, the polypeptide of claim 22, or any combination thereof.
 24. The cell of claim 23, wherein the cell is a T cell.
 25. The cell of claim 24, wherein the T cell is an allogeneic T cell, an autologous T cell, an engineered autologous T cell (eACT), or a tumor-infiltrating lymphocyte (TIL).
 26. The cell of claim 24 or 25, wherein the T cell is a CD4+ T cell.
 27. The cell of claim 24 or 25, wherein the T cell is a CD8+ T cell.
 28. The cell of claim 24 or 25, wherein the T cell is an in vitro cell.
 29. The cell of claim 24 or 25, wherein the T cell is an autologous T cell.
 30. A composition comprising the polynucleotide of any one of claims 1 to 19, the vector of claim 20 or 21, the polypeptide of claim 22, or the cell of any one of claims 23 to
 29. 31. The composition of claim 30, which is formulated to be delivered to a subject, optionally, comprising at least one pharmaceutically-acceptable excipient.
 32. A method of making a cell expressing a CAR or TCR comprising transducing a cell with the polynucleotide of any one of claims 1 to 19 under suitable conditions.
 33. The method of claim 32, further comprising isolating the cell.
 34. A method of inducing an immunity against a tumor comprising administering to a subject an effective amount of a cell comprising the polynucleotide of any one of claims 1 to 19, the vector of claim 20 or 21, the polypeptide of claim 22, or any combination thereof.
 35. Use of the polynucleotide of any one of claims 1 to 19, the vector of claim 20 or 21, the polypeptide of claim 22, the cell of any one of claims 23 to 29, or the composition of claim 30 or 31 for the manufacture of a medicament for treating a cancer in a subject in need thereof.
 36. The use of claim 35, wherein the cancer is a hematologic cancer.
 37. The use of claim 35, wherein the cancer is of the white blood cells.
 38. The use of claim 35, wherein the cancer is of the plasma cells.
 39. The use of any one of claims 35 to 38, wherein the cancer is leukemia, lymphoma, or myeloma.
 40. The use of any one of claims 35 to 38, wherein the cancer is acute lymphoblastic leukemia (ALL) (including non T cell ALL), acute myeloid leukemia, B cell prolymphocytic leukemia, B-cell acute lymphoid leukemia (“BALL”), blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloid leukemia, chronic or acute leukemia, diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), hairy cell leukemia, Hodgkin's Disease, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, Marginal zone lymphoma, monoclonal gammapathy of undetermined significance (MGUS), multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma (NHL), plasma cell proliferative disorder (including asymptomatic myeloma (smoldering multiple myeloma or indolent myeloma), plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, plasmacytomas (including plasma cell dyscrasia; solitary myeloma; solitary plasmacytoma; extramedullary plasmacytoma; and multiple plasmacytoma), POEMS syndrome (also known as Crow-Fukase syndrome; Takatsuki disease; and PEP syndrome), primary mediastinal large B cell lymphoma (PMBC), small cell- or a large cell-follicular lymphoma, splenic marginal zone lymphoma (SMZL), systemic amyloid light chain amyloidosis, T-cell acute lymphoid leukemia (“TALL”), T-cell lymphoma, transformed follicular lymphoma, or Waldenstrom macroglobulinemia, or a combination thereof. 